The nurse recalls that hemolytic anemia can be caused by which extrinsic factors

Asian J Transfus Sci. 2019 May; 13(Suppl 1): S32–S100.

2019 May; 13(Suppl 1): S32–S100.

PP 01: Clinical outcome of platelets transfusions using platelet-rich plasma platelets and buffy coat removed platelets in patients with thrombocytopenia

Background: Platelet transfusions are widely used to treat thrombocytopenia of various etiology. There are two different methods of preparation of platelet concentrate from whole blood, one is Platelet-rich plasma method (PRP) and the Buffy Coat Removed (BCR) method. A large number of studies have compared in-vitro activation of platelets among BCR platelets and PRP platelets but their significant clinical effects in patients has not been evaluated in any of these studies.

Aim: Aim of our study is to compare clinical outcome of patients transfused with platelets prepared by PRP method and BCR method.

Materials and Methods: This study was conducted in Dept of IHBT, JMMC Thrissur, over a period of two years. A total of 100 patients with thrombocytopenia were enrolled into the study with fifty patients in each group. Outcome of patients transfused with PRP derived platelets and Buffy coat removed platelets were compared on the basis of absolute and corrected count increment, percent platelet recovery and incidence of post transfusion reactions.

Results: The mean absolute count increment in BCR group was 23,900/μl with a standard deviation of 7022.56/μl. The mean absolute count increment in PRP group was 18,910/μl with a standard deviation of 7482.42/μl. The difference is statistically significant with a p value of 0.001. The mean CCI of PRP group is 12847 with a standard deviation of 5146.76 and in BCR group it is 12897 with a standard deviation of 4266.82 and this difference was not statistically significant p value (0.957). None of the patient transfused with BCR platelets reported a transfusion reaction while one out of 50 (2%) patients transfused with PRP platelets had FNHTR.

Conclusion: On the basis of count increment, corrected count increment, percent platelet recovery and incidence of post transfusion reactions, BCR platelet transfusions showed a better outcome than PRP platelet transfusions.

2019 May; 13(Suppl 1): S32–S100.

PP 02: Therapeutic efficacy of plasma exchange in the treatment of myasthenia gravis

Background: Myasthenia gravis (MG) is a well known autoimmune disease characterized by antibodies against the acetylcholine receptor (anti-ACHR) on the post synaptic surface of the motor end plate. Plasma exchange is a therapeutic modality well established in MG with a positive recommendation based on strong consensus of class III evidence and in the category 1.

Aims: The aim of this study was to analyze the retrospective experience related to the indication, complication and outcome of Therapeutic Plasma Exchange (TPE) in Myasthenia Gravis.

Methods: A total of 35 patients of MG were submitted to a total of 41 cycles and 171 sessions of TPE. It was performed using a single volume plasma exchange with intermittent cell separator (Haemonetics MCS plus) by femoral or central line access and scheduled preferably on alternate-day intervals for eight to ten days. Both subjective and objective clinical response to TPE was estimated and final assessment of response was made at the time of the last TPE in the series and overall outcome at discharge.

Results: Total of 110 patients of MG who were admitted to our hospital during the study period of 2 years. 35 (31.8%) patients had TPE performed with mean age of 32 years (M: F=2:1). The mean number of TPE session was 4.2 (SD±1.2), volume exchange was 2215 ml (SD±435), overall incidence of adverse reaction was 21.7%. All patients had immediate benefits of each TPE cycle. Good acceptance of procedure was observed in 78.3% of patients.

Conclusion: TPE may be considered as one of the treatment options especially in developing countries like ours as it is relatively less costly but as effective for myasthenic crisis as other modalities. It may be more effective if initiated earlier in the hospital course and in patients who had previously failed to respond to other treatment.

2019 May; 13(Suppl 1): S32–S100.

PP 03: Role of therapeutic plasma exchange in autoimmune haemolytic anaemia - A case report

Background: Autoimmune Haemolytic Anaemia (AIHA) represents a group of disorders in which autoantibodies mediate either intravascular haemolysis by the terminal lytic complex (C5b-C9) or, more often, extravascular destruction in the spleen by the macrophage-phagocytic system. AIHA can be classified into two major types, Warm Autoimmune Haemolytic Anaemia (WAIHA) and Cold Agglutinin Disease (CAD)/Cold Autoimmune Haemolytic Anaemia (CAIHA). In WAIHA, the Direct Antiglobulin Test (DAT) is positive with Anti-IgG. In CAIHA, the DAT is positive with Anti C3b only. Therapeutic Plasma Exchange (TPE) is typically utilized in patients with fulminant haemolysis who are unresponsive to Red Blood Cell (RBC) transfusion. TPE treatment may temper the disease course until immunosuppressive therapy takes effect, or if other treatments have failed.

Case Report: We report two cases, one of WAIHA and other of CAIHA, in which TPE was done to manage the patient. In case of WAIHA, six daily cycles of TPE were performed and in case of CAIHA, one cycle of TPE was performed. In both cases, TPE was successful in removing autoantibodies and there was significant increase in haemoglobin level post transfusion.

2019 May; 13(Suppl 1): S32–S100.

PP 04: Cytokines in investigation of blood transfusion reactions

Background: Atleast 3% of all transfusions result in either a febrile non-hemolytic transfusion reaction (FNHTR) or allergic reaction. FNHTRs are seen in 1% to 3% of RBC transfusions. The mechanism of FNHTR involves antigen antibody complex, complement activation and release of cytokines like tumor necrosis factor Interleukin (IL)-1, IL- 6, IL-8 which have pyrogenic effect. Regulated upon activation, normal T cells expressed and secreted (RANTES) is mainly involved in allergic reactions.

Aims: To investigate blood transfusion reactions for cytokine levels.

Methods: Routine investigation protocol of the blood bank was carried out for reported blood transfusion reactions (BTR). When no root cause of reactions was found nine cases were further investigated for cytokines The pre and post sample of the patient and the donor's sample from the residual bag were investigated for IL-6, IL-8, IL-1β and RANTES using ELISA.

Results: The selected patients were of age group 18 to 58 years. Three were male and six female. Three patients were transfused with whole blood and six with red cell concentrate. In whole blood transfusion IL-6 doubled from 101 to 201 pg/ml in post transfusion sample whereas it was absent in the bag, IL-8 increased from 54 pg/ml to 894 pg/ml and IL-1β was 60pg/ml which was absent in pre sample. In RCC transfusion RANTES was absent in the patient but it was 98ng/ml and 131 ng/ml in blood bag. IL-8 and RANTES was found in high levels in all the bags.

Conclusion: RANTES accumulates in large amount in blood components. It may be possible that when these are transfused to patients it may be possible to cause allergic symptoms that are associated with NHTR. IL-6 and IL-8 also increased which may be the cause of febrile reactions due to cytokines released from leucocytes. Thus investigation of BTR can be extended from routine compatibility testing.

2019 May; 13(Suppl 1): S32–S100.

PP 05: Study of exchange transfusion by reconstituted blood in hemolytic disease of newborn

Background: This study was aimed to review and establish the practice of exachange transfusion with reconstituted blood in neonates and to observe fall of bilirubin and its comparison with related studies.

Aims: (1) To find out fall of indirect bilirubin level after exchange transfusion in newborn. (2) To establish the role of reconstituted blood for exchange transfusion in newborn.

Methods: 31 Neonates diagnosed hemolytic disease of fetus & newborn were selected for this study, in which exchange transfusion was carried out as one of the treatments for hyperbilirubinemia. Out of the 31 cases, 20 were of resus (rh) hemolytic disease of fetus and newborn, while abo and other blood groups constituted 8 and 3 hemolytic disease of fetus and newborn cases respectively. First, the neonates’ and mother's blood samples were subjected to relevant investigations. After that, for neonates having rh hemolytic disease of fetus and newborn, o rh negative cells suspended in ab plasma were given, o rh positive cells suspended in ab plasma were given to abo hemolytic disease of fetus and newborn because of other blood group antibodies. The exchange transfusion was carried out taking all aseptic precautions by continuous technique with double-volume exchange transfusion method.

Results: The average post-exchange fall in serum indirect bilirubin was (53.70%) in all 31 cases, which was found to be more significant that the previous studies. Looking into the superiority of the exchange transfusion in hemolytic disease of fetus and newborn by reconstituted blood, the reconstituted blood can be modified and suppleid as per the requirement and conditions.

Conclusion: From this study we concluded that reconstituted blood is immunologically safer & better than whole blood for purpose of exchange transfusion in hemolytic disease of fetus&newborn because of its superiority in minimizing transfusion reaction and in achieving all the therapeutic effects of exchange transfusion in better way.

2019 May; 13(Suppl 1): S32–S100.

PP 06: Study of usage of fresh frozen plasma and effect of fresh frozen plasma on pretransfusion international normalised ratio

Background: Fresh Frozen Plasma is frequently prescribed blood product used therapeutically in bleeding or prophylactically in non-bleeding patients prior to invasive procedures or surgery. High rate of inappropriate usage has been reported that leads to wastage of limited resources, depriving needy patients of their use. Also, there is a risk of transmission of infectious agents. This study was undertaken to audit the uses of FFP and to assess effect of FFP on pre transfusion INR.

Aim: To study the effect of FFP on pre transfusion INR, To audit usage of FFP.

Methods: During the study 500 patients were prospectively observed who received FFP in our hospital. FFP usage was classified as appropriate or inappropriate based on guidelines by the National Health and Medical Research Council. Pre and Post transfusion INR were recorded and effect of FFP on pre transfusion INR was studied in patients who appropriately received FFP.

Results: Total 2048 units were issued for the 500 patients of which 158 were females and 342 were males. (Range: 17-54 years). Total 1698 units in 415 patients were appropriately transfused and 350 units in 85 patients were inappropriately used. Mean improvement in pre-transfusion INR per unit of FFP was 0.26 (median 0.25, range 0.02 to 1.1, SD 0.16). A significant improvement in the pre transfusion INR per unit of FFP was seen in 60.7% patients. A linear relationship was noted between pre-transfusion INR and improvement in INR per unit of FFP.

Conclusion: Proportion of inappropriate FFP usage remains high. A significant improvement in INR is more likely with a high pre-transfusion INR. The improvement in INR per unit of FFP is also more with higher pre-transfusion INR. Awareness programs regarding blood component usage in various clinical conditions should be conducted for clinicians regularly.

2019 May; 13(Suppl 1): S32–S100.

PP 07: Single dose intra-articular platelet-rich plasma versus corticosteroid injections in the treatment of adhesive capsulitis of the shoulder: A cohort study

Background: Adhesive capsulitis (AC) is one of the common causes of shoulder pain and disability. Intra-articular corticosteroid (IA-CS) injection still remains one of the most common procedures for treating AC. However, corticosteroid injection has been associated with hyperglycaemia, detrimental effects on articular cartilage, increased risk of tendon rupture, local skin depigmentation and atrophy of subcutaneous tissue. Recently, new evidences have emerged on the effectiveness of Platelet Rich Plasma (PRP) injection in the treatment of chronic tendon and muscle injuries, tendinopathies, osteoarthritis etc. However, its evidence of effectiveness in patients with AC is limited.

Aims: The objective of this study was to compare the effects of single IA-PRP injection with conventional single IA-CS injection on pain, active and passive shoulder range of motion (ROM) in patients with primary AC.

Methods: This was prospective observational, cohort study. Patients, contraindicated to standard CS injection, received IA-PRP injection (n=30), whereas patients, with no contraindication, received standard IA-CS injection (n=30). In IA-PRP group, we administered ultrasound guided single intra-articular injection (4 mL) of PRP into the GH joint, whereas, in IA-CS group, we administered single injection of 2 mL (40 mg) of Methyl Prednisolone Acetate, mixed with 2 mL 2% lignocaine (Total 4 mL) into the GH joint. The PRP was prepared by the single centrifugation technique using bench top centrifuge System (Eppendorf AG Centrifuge 5702), in accordance with standard operating procedure.

Results: 28 patients in IA-PRP group and 27 patients in IA-CS group finished the entire 12 week study period. The improvement in pain intensity, shoulder ROM and shoulder function score was significantly greater in the IA-PRP injection group than in IA-CS injection group (p<0.05).

Conclusion: Single dose IA-PRP injection was found to be more effective than an IA-CS injection, in improving pain, disability, and shoulder ROM in patients with primary AC of the shoulder.

2019 May; 13(Suppl 1): S32–S100.

PP 08: Role of therapeutic plasma exchange in thrombotic thrombocytopenic purpura

Background: Thrombotic thrombocytopenic purpura (TTP) is a rare and life-threatening thrombotic microangiopathy characterized by microangiopathic hemolytic anemia, severe thrombocytopenia, and organ ischemia linked to disseminated microvascular platelet rich-thrombi. This disorder has feature of antibody production resulting in endothelial damage and/or deficiency of ADAMTS-13. Therapeutic plasma exchange (TPE) comprises of withdrawing offending agent in patient's plasma causing clinical symptoms and replacing with either donor plasma or manufactured replacement solution such as 5% albumin and 0.9% normal saline.

Aim: To observe the clinical benefits of TPE in a patient with thrombotic thrombocytopenic purpura.

Methodology: This study was carried out in department of Immuno Hematology and Blood Transfusion, Ramachandra Medical College and Research Institute, Porur, Chennai. Continuous flow cell separator (COMTEC) was used for TPE. Effect of TPE was observed in a patient with TTP. Total 6 sessions of TPE was done for a female middle aged patient in 6 consequent days in the study period of May 2018 to June 2018. Patient had complaints of left hand numbness and slurring of speech, deviation of angle of mouth and blurring of vision since 10 days. Femoral catheter was inserted in that patient before procedure. At each procedure 1-1.5 plasma volume exchange was done and FFP in combination with 0.9% normal saline and 5% albumin were used as replacement fluid.

Results: Total 6 sessions of TPE performed. She showed clinical improvement and her lab para-meters were reviewed daily. Her platelet count improved drastically from 15,000 to 2,38,000/mm3, LDH had fallen from 730 to 228U/L, schiostocytes from 8% to 0.5% by the end of 6th cycle. Complications such as Hypocalcemia (perioral tingling), hypomagnesia (muscle cramps), chills and rigor and rash were recorded during procedure and were addressed periodically. No fatal complications were observed. Patient recovered uneventfully and was discharged.

Conclusion: TPE is an ideal and first line treatment (category I indication for TPE) option in patients suffering from TTP with minimal complications and better outcome.

2019 May; 13(Suppl 1): S32–S100.

PP 09: Transfusion support in trauma induced coagulopathy

Introduction: Blood transfusion support is vital for patients of trauma with haemorrhagic shock. Bleeding remains the leading cause of preventable death after injury. Coagulopathy after traumatic injury is multifactorial and involves all components of hemostatic system.

Case Discussion: A Case Of 28 Years Old Male Presented To The Casualty With Bluntinjury abdomen of heavy object on his abdomen. On examination – Patient was conscious, coherent, cooperative GCS-15/15. Vitals –pulse-130/min BP-80/60/mm Hg RR-24/min. On USG –haemoperitoneum, splenic and hepatic contusions seen. Investigations – Hb-10gm/dl, TLC-21000/cumm, PLT-1.4lakshs/cumm. Patient taken for exploratory laparotomy. Transfusion support -1 pint PRBC and 1 unit FFP given prior to surgery intra op -2 pints of whole blood and 1 unit FFP given with damage control surgery. Postoperative findings – POD-1 - INR-2.14 PT-27 Seconds. In view of raised INR-2 FFP were given

• POD-2-Hb-dropped to 7 gm/dl so 2 PRBc were given

• POD-3 - Hb-8.3gm/dl PLT-50000cumm INR-1.8.

Smear-microcytic hypochromic anemia with thrombocytopenia, one unit of single donor platelets and 1 FFP was given Next Day, reexploratory laparotomy was done for pack removal and no active bleed was found. On post operative findings Hb- 9.4 gm/dl –one unit of PRBC was given. Patient clinically improved and got discharged in a haemodynamically stable condition. Treatment – Iv Fluids, Antibioticsand Analgesics - given during the st.

Dicscussion: Blood administration during truncal surgery with uncontrolled bleeding includes infusion with compression devices, high volume rapid infusion pumps, blood salvage system provides rate of flow at even 100 m/min.

Conclusion: (1) End goal of transfusion is to restore volume and oxygen carrying capacity. (2) Standard coagulation tests and functional viscoelastic assays are used for diagnosis. (3) Balanced resuscitation and optimal monitoring is the mainstay for trauma induced coagulopathy

2019 May; 13(Suppl 1): S32–S100.

PP 10: Platelet count kinetics in dengue fever

Introduction: Dengue fever ranges from asymptomatic infection to severe dengue shock syndrome. Thrombocytopenia in Dengue has been an universal finding. Platelet count kinetic studies demonstrate a fall in platelet count from Day 3 of fever through day 7 and then rise to normal levels from day 8.

Aim: To study the platelet count kinetics in Dengue patients.

Methods: Case records of 110 patients admitted with Dengue were analyzed for clinical, laboratory parameters and transfusion requirements during hospitalization.

Results: Ten pediatric patients and 100 adults were included in the study. The median age of the pediatric patients was 7 years. The mean platelet count at admission was 0.57 ±0.37 lakhs/μl. The lowest platelet count seen was on median day 5 of fever and the mean lowest platelet count was 0.16±0.06 lakhs μl. Platelet count returned to >50,000 on median day 9 and platelet count increased to >1,00,000 lakhs/μl on median day 10. Median Hospitalization days were 10. 7/10 pediatric cases were severe dengue and remaining were dengue with warning signs. Mortality was 30%. Adults: Patients were divided in to Non-Transfused (n=46) and Transfused Group (n=54). The mean platelet count at admission was 0.36 lakhs/μl in transfused and 0.76 lakhs/μl in the non-transfused group. The lowest platelet count observed in the non-transfused group was 0.27 lakhs/μl and 0.13 lakhs/μl in transfused group on day 7. Platelet count recovered to >50,000/μl on day 9 and to >1,00,000/μl on day 10 in both the groups.

Conclusion: Platelet count recovery to hemostatic level was observed on day 8 to day 10 in majority of the cases. No difference in platelet count kinetics could be observed between transfused and non-transfused group. No significant difference in outcomes like complications and hospital stay was observed between transfused and non-transfused patients.

2019 May; 13(Suppl 1): S32–S100.

PP 11: Autologus blood transfusion: An experience in a tertiary care hospital of NCT Delhi

Introduction: Autologus Blood Transfusion (ABT) is the safest mode of blood transfusion as it eliminates risk of transfusion transmitted infections, allo-immunisation and graft versus host disease. ABT includes replacement of blood loss during surgeryby patient's own blood donated before surgery or perioperatively. It is of immense help in case of rare blood group patients and in maintaining blood transfusionservices in remote areas.

Aims and Objective: (1) To assess the efficacy of ABT. (2) To study the effect of perioperative ANH on the haematological and coagulation parameters.

Materials and Methods: Study of autologous blood transfusion casesover a period of two years. The patients were selected for ABT as per national guidelines. Single unit of 350 ml was collected from each patient in pre-surgical donations and approx. 10 ml/kg of blood was withdrawn in perioperative hemodilution. The details of all the patients for autologous blood transfusion were recorded.

Results and Observations: A total of 102 (0.17%) cases reported for ABT out of 61000 of total donation in two years. The various surgical procedures which patients underwent included: Cholecystectomy (37), Hernial repair (25), urolithotomy (12), non healing fractures (20) and vaginal hysterectomy (08). 55 units were issued asautologous BT for same patients, 40 units were cross over to main stock as transfusion in these patients were not required. Three units were expired due to inventory control failure and two were found reactive. Perioperative ANH was done in 25 cases. ANH showed dilutional effect on coagulation and haematological parameters but PT & PTTK returned to near baseline valueson the 2nd post op day, however values of haematological parameters did not revert back to baseline.

Conclusions: ABT is used infrequently in developing countries despite the fact that it is free of risks associated with allogenic transfusion, helps in conserving blood stock and provision of blood to rare blood group patients.

2019 May; 13(Suppl 1): S32–S100.

PP 12: Nature and causes of errors in the blood transfusion chain - A step towards patient safety

Background: A wide range of errors (all deviations from Standard Operating Procedures) occur at various steps of the transfusion process.

Aim: This study was conducted to identify the nature and cause of errors in the process of blood transfusion.

Materials and Methods: A prospective study was conducted in the department of Transfusion Medicine at a tertiary care hospital in India over a period of 18 months. All the errors that occurred during the process of blood transfusion starting from the donor phlebotomy till the transfusion of blood component to the patient were reported and analyzed.

Results: The rate of error occurrence was found to be 0.3%. The actual events which include near-harm and adverse events were 10% and 5%, respectively whereas near-miss events were 85% (ratio of actual to near-miss was 1:5.6). Laboratories were found to have the highest level of error occurrence (48% of all errors). Wrong blood in tube errors were found to be 1 per 3308 samples received. Frequency of haemolytic transfusion reactions due to ABO incompatibility was found to be 1 in 6770. Majority (74.4%) of the errors were recovered due to system check built within the departmental SOPs thus, recovery was planned. However, the remaining 25.6% events were discovered only after the product was issued.

Conclusion: The data obtained through this study highlighted various critical steps where errors can occur in the long chain of blood transfusion.

2019 May; 13(Suppl 1): S32–S100.

PP 13: Analysis of demographic and clinical variables on influencing blood loss and transfusion requirement in unilateral total hip arthroplasty

Background: Many advances in surgical and anaesthetic techniques on total joint arthroplasty have taken place in last few decades, still there is associated with considerable peri-operative blood loss during this procedure especially in total hip arthroplasty (THR).

Aims: The Main Purpose Of This Study Was To Identify The Influence Of Different Demographic and clinical variables on perioperative blood loss and transfusion requirement especially in terms of age, sex, BMI, aetiological factors of the disease conditions and pre-operative haemoglobin (HB) in patients undergoing THR.

Methods: This was a retrograde observational study. Patients undergoing unilateral THR from January 2016 till July 2017 meeting inclusion criteria were included in our study. Peri-operative blood loss was calculated with use of validated formula. Blood transfusion requirement was also evaluated with respect to other variables.

Results: Total 81 patients were included for retrospective analysis in our study. Mean patient age was 43.71± 15.03 years, mean BMI was 26.4±4.1, mean pre-op HB was 11.25±1.57 gm/dl, mean blood loss was 361.42±295.94 ml and median blood transfusion requirement was 01 (IQR:1-2). Patients were broadly categorised into three major groups [#Neck Of Femur (NOF), N=17, Inflammatory, N=35 and Avascular necrosis hip, N=29] on the basis of aetiology of the disease conditions. NOF group had highest and significant blood loss (271.17±71.92 ml, P<0.001) compared to the other two groups. Further regression analysis showed that etiological factors, sex, and pre-op HB had a significant influence on blood loss, whereas aetiological factors and sex had an influence on transfusion requirement (P<0.05). Patient with lower HB (HB<12 gm/dl) showed significant negative correlation with transfusion requirement (P<0.05).

Conclusion: Our study identified three different significant variables which influence blood loss and transfusion requirement in patients undergoing THR which we believe can be used for implementing more effective blood conservation strategies.

2019 May; 13(Suppl 1): S32–S100.

PP 14: Plasmapheresis as an adjuvant in the treatment of antibody mediated rejection

Introduction: Antibodies are known to cause rejection of the transplanted organ. Plasmapheresis (TPE) offers therapeutic benefit by clearing these antibodies from circulation. We carried out retrospective analysis of antibody mediated rejections (AMR) in renal transplant recipients and evaluated the therapeutic efficacy of plasmapheresis.

Methods: This is a study of set of patients. We reviewed case files of patients who underwent TPE in our department from January 2017 till December 2017. Patients with biopsy proven AMR were subjected to TPE (Group-1) were considered for study. Serum creatinine (SCr) levels before and after TPE were evaluated and compared with controls who did not undergo TPE (Group-2). On an average three cycles of plasmapheresis were carried out either on Spectra Optia and Fresenius Kabi Comtech apheresis system. Approximately 1 to 1.5 times plasma volume was removed per cycle and replacement was carried out with crystalloids and colloids in ratio of 3: 2 (0.9% saline and 20% human albumin).

Results: Out of the 156 patients showing T+B cell or B cell rejection. 24 patients opted to undergo TPE (Group 1) and 132 patients refused to undergo TPE. The mean age of the patients in group 1 was 34.5 years M: F ratio was 6:1 and mean SCr before and after TPE was 3.40 mg/dL (S.D 1.68) & 2.34 mg/dl (S.D 1.20) respectively. The mean age of the patients in group 2 was 35 years, M: F 9.2:1 and mean SCr at the time of diagnosis and last follow up was 2.89 mg/dl (S.D 1.66) and 3.28 mg/dl (S.D 1.87) respectively.

Conclusion: Plasmapheresis is a useful adjuvant of anti-rejection therapy for AMR.

2019 May; 13(Suppl 1): S32–S100.

PP 15: Vascular access and access related complications in therapeutic plasmapheresis among patients of neurological disorders: A tertiary care centre experience

Background: Therapeutic plasma exchange (TPE) is an apheresis modality in which plasma is separated from the blood, discarded, and replaced with an isosmotic fluid. One of the critical aspects for the successful performance of TPE is appropriate vascular access to provide high blood flow for the collection and return phases of the procedure, because most patients undergoing TPE will require more than one treatment over days.

Aims: To study access used for performing TPE in neurology patient and analyze access related complications.

Methods: A prospective collection of data of TPE was done for 4 months. All consecutive patients who underwent TPE were included. The demographic information, diagnosis, access used, TPE procedure details and access related complications were recorded. All the TPE procedures were done on MCS+ cell separator.

Results: A total of 221 including 123 male and 99 female patients underwent TPE and 999 cycles were performed. The average age was 39 (range 11- 74) years. The majority of patients were of Guillain-barré syndrome (98/221, 44.34%), Myesthenia Gravis (n=28,12.67%), Neuromyelitis Optica- Spectum disorder (n=27,12.22%) and autoimmune encephalitis ((n=27,12.22%). The access used were peripheral antecubital in 178 (80.54%), Central venous catheter (CVCs) in 20 (9.05%) and a combination of both peripheral and CVCs in 23 (10.4%) patients. Among the last group initial cycles were performed by peripheral access and subsequently CVCs were used. The hematoma (n=20) and phlebotomy failure (n=20) for peripheral access related and blocked central line (n=4) for CVCs were most common complications.

Conclusion: The peripheral venous access is preferred method of TPE access at our centre. This is contrary to international apheresis survey of 2010 which states marked shift to CVCs in Asian countries. This mainly may be due to non-participation of many Asian countries. It is preferred options for TPE in most of European countries but CVCs are preferred in American countries.

2019 May; 13(Suppl 1): S32–S100.

PP 16: Atypical hemolytic uremic syndrome: Single center experience of therapeutic plasma exchange

Introduction: Hemolytic uremic syndrome is a disease characterized by triad of hemolytic anemia, acute kidney failure and a low platelet count. HUS is usually categorized as typical, caused by Shiga toxin–producing Escherichia coli (STEC) infection, as atypical HUS (aHUS), usually caused by uncontrolled complement activation, or as secondary HUS with a coexisting disease. Plasma therapy is the mainstay of treatment as the drug Eculizumab is not available in India. We did a retrospective analysis of the patients who underwent Therapeutic plasma exchange as a treatment for atypical HUS.

Materials and Methods: A retrospective analysis of the patients who were referred to the department of Transfusion Medicine for Therapeutic plasma exchange was done from the year January 2017 to June 2018. All patients with clinical diagnosis of atypical HUS referred by the department of Pediatric Nephrology for Therapeutic Plasma Exchange between 2017 to June 2018 were included in the study. Volume of plasma removed was decided according to the clinical condition of the patient by the consulting doctor. Fresh Frozen Plasma was used as a replacement fluid in all the cases.

Results: Fifteen patients were referred for Therapeutic Plasma Exchange. Five patients were Females and 10 were males. Age of the patients was from 4 years to 11 years with a mean age of 6 years. A total of 119 plasma exchanges were performed for these 15 patients. Average pre procedure platelet count and Hemoglobin was 96533 per cubic millimeter and 6.76 gm% respectively. Average platelet count and Hemoglobin at the end of last procedure was 1,16,800 per cubic millimeter and 8.1%. None of the patients developed adverse reaction during the procedure.

Conclusion: Therapeutic Plasma Exchange is a safe procedure and is an adjuvant in the treatment of atypical HUS.

2019 May; 13(Suppl 1): S32–S100.

PP 17: An audit of usage of platelets in dengue cases in a tertiary care hospital

Introduction: Judicious use of platelets in dengue cases needs to be done to avoid unnecessary adverse transfusion complications and better management of platelet inventory.

Aim: To audit if appropriate utilization of platelets is done in dengue patients.

Materials and Methods: The study was done from July 2015 to June 2018. Seropositive dengue cases were included in the study. BMC- EPID cell and DGHS guidelines were used as reference (Prophylactic platelets may be given in dengue at level of <10,000/cumm in absence of bleeding manifestation). A written notice was circulated to the clinicians/wards/ICUs regarding the BMC- EPID cell protocol and a format was prepared for documenting the lab details of patient (platelet count, PT, APTT, HCT, Indication). If count of any patient was more than 10,000, history of bleeding manifestation was elicited and correlation with IPF (Immature platelet function) was also done. If no significant history was found, the platelets were reserved for the patient and the clinician was requested to further monitor the patient and platelet count and avoid platelet transfusion if possible. All the requests for platelets for dengue cases were analysed.

Results: Among the total 832 cases of dengue positive patients, requests for platelets for 38 (4.5%) patients had been received (7 had platelet count <10,000; 9 between 11000-20000 and 22 between 21,000 to 40,000). Out of the request received, 22 (2.6%) patients required platelet transfusion - 7 had platelet count <10,000; 10 had some bleeding manifestation and 5 had count between 11,000 to 20,000/cumm. These were transfused on the physician's clinical judgment. All the platelet transfusion were justified as per the guidelines.

Conclusion: Every blood bank needs to implement guidelines and monitor appropriate use of platelets in dengue patients. Proper coordination between clinician and blood bank plays an important role in promoting rational use of platelets.

2019 May; 13(Suppl 1): S32–S100.

PP 18: Hemovigilance - An analysis of transfusion reactions among recipients in a tertiary-care centre through an active self surveillance system

Introduction: Hemovigilance is defined as a set of surveillance procedures covering the whole transfusion chain intended to collect and assess information on unexpected or undesirable effects resulting from the therapeutic use of labile blood products and to prevent occurrence or recurrence of such incidents. We as a tertiary care centre studied the entire blood transfusion reactions among the patients over a period of six months. Initial three months includes the period where clinicians reported all those transfusion reactions which they found as definite cases which can be attributed to transfusion. Last three months includes the period which we adopted an active self surveillance to trace back all blood components which are transfused and imparting awareness among the clinicians about necessity of transfusion reaction reporting.

Aims and Objectives: To analyze the incidence of adverse transfusion reactions, to improve the adverse reaction reporting system to maximize patient safety, to develop strategies to reduce incidence of transfusion reactions.

Results: The Reporting rate for Transfusion reaction has significantly increased during the last 3 months (8 reactions out of 1637 transfusion) where self surveillance was adopted as a method of active hemovigilance compared to first 3 months (2 reactions out of 1536 transfusions). We could trace back all components which were transfused over a period of six months and reactions produced by the various components. Previously only definite reactions were reported by the clinicians. Now, since the clinicians are more aware about the hemovigilance reporting system due to the self surveillance, more number of cases are being reported. Now, we could classify all the reported cases into various imputability categories as described by the recipient vigilance system.

Conclusion: Active hemovigilance will help us to improve the patient safety by adopting safe transfusion practice with improved quality of blood components and by increasing awareness among clinicians.

2019 May; 13(Suppl 1): S32–S100.

PP 19: Peripartal blood products transfusions in association with hellp syndrome and preeclampsia

Aims: (1) To assess blood product requirement during peripartal period in preeclampsia and HELLP syndrome. (2) To assess the outcome of blood products transfusion in Preeclampsia and HELLP syndrome.

Methods: Retrospective, descriptive study on 20 peripartal mothers with either Preeclampsia or HELLP syndrome, which were reviewed with special attention to laboratory data, details of transfusion therapy.

Results: We reviewed 20 peripartal women from june 2017- august 2018 period, out of which 15 cases were preeclampsia and 5 were having HELLP syndrome. Among 15 cases of preeclampsia only 2 required blood products (13.3%), whereas all 5 peripartal mothers with HELLP syndrome requried blood products (100%). Preeclampsia mothers required blood products in form of PRBC and FFP, whereas in mothers with HELLP syndrome required additional SDP along with PRBC and FFP (3 in 5 reqired prbc and FFP, whereas all 5 requird SDP). Post transfusion Hemoglobin in 2 cases of preeclampsia which were transfused with prbc was maintained in a range of 9 - 10 gm/dl (100%), whereas in HELLP syndrome out of 3 transfused with prbc, only one case maintained hemoglobin at a range of 9-10 gm/dl (33.3%), other two cases maintained hemoglobin at 7-9 gm/dl (66.6%). Post transfusion PT, aPTT, INR were maintained in normal range in 2 cases of preeclampsia, who were transfused with FFP, whereas in mothers with HELLP syndrome all 3 cases didnot maintain PT, aPTT, INR in normal range despite FFP transfusion. Post transfusion platelet count in 4 of 5 transfused with SDP in HELLP syndrome maintained >50000cells/cumm (80%), one maintained <50000 cells/cumm (20%).

Conclusion: Blood products requirement was more in HELLP syndrome compared to preeclampia. SDP requirement was seen in HELLP syndrome. Post transfusion levels of hemoglobin, PT, aPTT, INR were maintained in the normal range in preeclampsia.

2019 May; 13(Suppl 1): S32–S100.

PP 20: Clinical outcome of therapeutic plasma exchange in two oncology patients of different age groups with guillain barre syndrome

Background: Guillain Barre Syndrome (GBS) is anautoimmune condition characterized by progressive ascending paralysis and areflexia with or without abnormal sensory and autonomic functions that reaches maximum severity in four weeks of onset of symptoms. GBS is a category 1 indication for therapeutic plasma exchange according to the ASFA guidelines.

Aim: To understand the clinical course, severity and outcomes in two different oncology cases with superadded GBS where plasma exchange was started within four weeks of onset of symptoms.

Methods: Two different cases of GBS were followed, one pediatric patient with Pre B-ALL who was IVIG resistant and one adult patient with adenocarcinoma stomach as the primary diagnosis who was on ventilator support. Both the patients were started on IVIG at some point of their hospital stay, however because of below par clinical improvement, therapeutic plasma exchange (TPE) was considered. Five procedures of TPE were performed on both of these patients. Approximately 1 plasma volume was replaced in the pediatric patient in each sitting and 1.5 plasma volume in the adult patient.

Results: The pediatric patient showed significant increase in power in the upper limbs by the end of the 3rd sitting of TPE but no significant improvement was seen in the lower limbs. The adult patient however showed greater response and jumped two points on the GBS disability score from 5 to 3 and he was able to walk with assistance within 2 weeks of the procedure.

Conclusion: TPE should be considered early on, to arrest the disease progression as well as for quicker recovery. GBS is most responsive to plasma exchange early on, after the onset (<2-4 weeks). Disease progression and clinical outcomes of GBS are highly variable but most patients begin to recover around 28 days following onset.

2019 May; 13(Suppl 1): S32–S100.

PP 21: Granulocyte transfusions in hemato-oncology patients: One-year experience from a tertiary care oncology centre

Background: Hemato-oncology patients, who have received intensive chemotherapy, may develop severe neutropenia and serious bacterial and/or fungal infections. It is still under debate whether granulocyte transfusions (GTs) increase survival in patients with febrile neutropenia.

Aim: To evaluate clinical efficacy of granulocyte transfusions in Hemato-oncology patients with febrile neutropenia.

Materials and Methods: A retrospective analysis from January 2017 to December 2017 was done to evaluate clinical efficacy of nineteen granulocyte transfusions in fifteen hemato-oncology patients. Mobilization of the granulocyte donors was done with as per standard protocol.

Results: Minimum granulocyte yield of 1 X1010/bag was considered adequate for transfusion which was fulfilled in 90% of granulocyte harvest conducted. Over the entire study period, eligibility criteria for granulocyte transfusions were fever, an absolute neutrophil count (ANC) <500/μL, evidence of bacterial and or fungal infections (i.e clinical signs of infection, positive cultures and radiological evidence) and unresponsiveness to appropriate antimicrobial therapy for atleast 48 hours. Patients were grouped according to doses of granulocyte transfusions during the infectious episode (standard dose: 1.5-3.0 X 108 cells/kg and high dose: >3.0 X 108 cells/kg). Effect of clinical, microbiological and GT-related variables on infection-related mortality (IRM) was investigated. The post transfusion absolute neutrophil count (within 24 hours) increased significantly (median value: 350/μL) as compared to baseline levels (median value: 40/μL) (p<0.05). IRM was 33% in standard dose group and 8% in high dose group.

Conclusion: The absolute neutrophil count post transfusion showed significant increase as compared to baseline value. Granulocyte transfusions can be a valuable tool to improve outcome of infections in neutropenic patients provided adequate doses are transfused.

2019 May; 13(Suppl 1): S32–S100.

PP 22: The other “O”: Parabombay

Background: The Para-Bombay phenotype (also known as H-deficient secretor), is characterized by a lack of ABH antigens on the red blood cells, but presence of ABH substances in saliva. Bombay phenotype (H-deficient, non-secretor) is characterized by the absence of ABH blood group antigens both on the surface of red blood cells (RBCs) and in secretions. Both the Bombay and Para-Bombay phenotypes are the result of point mutations in the FUT1gene, which results in the formation of complete H-deficient (Bombay) or partial H-deficient (Para-Bombay) phenotype.

Aims: To present a case of a 40 year old patient admitted to our hospital requiring two units of blood, diagnostic workup and clinical implications.

Methods: Standard serologic techniques were used. Forward and Reverse grouping was done using Column Agglutination Technology (CAT), Testing with anti-H lectin, Indirect Antiglobulin Test (IAT), Compatibility testing by CAT done in our lab. Secretor studies, Adsorption and Elution were done in Reference Lab.

Results: Forward and Reverse grouping revealed the group to be ‘O’ positive. Anti-H lectin testing was negative. Indirect Coomb's test was negative. Secretor studies positive for presence of H substances in saliva. Adsorption and elution failed to demonstrate the presence of weak antigens on the RBCs.

Conclusion: Our serological tests were in line with the characteristics of para-Bombay phenotype. Problems may arise in finding compatible units for these patients because of anti-H or anti-IH, but most often these are not clinically significant. When Bombay or Parabombay blood is not available, whole blood units of normal ABO blood groups compatible by IAT are transfused, and the survival is expected to be almost normal. Two units of O positive blood were compatible for this patient, but unfortunately, our patient was discharged prior to transfusion and lost to follow up.

2019 May; 13(Suppl 1): S32–S100.

PP 23: Graft and patient outcome in abo-incompatible solid organ transplantation after desensitization with immuno-adsorption plasmapheresis

Background/Aims: Conventional and cascade plasmapheresis are commonly performed as part of desensitization protocol for ABO-incompatible solid organ transplants (SOT) and there are published reports from India. Recent introduction of isoagglutinin specific Immuno-Adsorption (IA) plasmapheresis offers certain advantages including processing of larger plasma volumes, quicker reduction of isoagglutinin titers and no requirement of replacement fluids. Since there is no published report on use of IA, authors’ center aimed to evaluate success rate of desensitization protocol, graft and patient outcomes when IA procedures were performed for desensitization in ABO-incompatible SOT.

Methods: The evaluation was done at large tertiary care center in north India. Patient records for preceding two-years were collated from hospital information system (HIS) and manual therapeutic apheresis procedure forms. Patients undergoing ABO-incompatible SOT with use of IA to decrease isoagglutinin titer were included in the study.

Results: During study period, 16 IA procedures were performed in total six patients who underwent successful ABO-incompatible SOT (4 kidneys/2 livers). The pre-IA isoagglutinin IgG titer ranged from 32-512. Mean number of IA procedures performed to achieve the desired pre-transplant IgG titer ≤8 was 2.66 (1-4). New IA column was used for each patient (and re-used for the same patient, if needed, after sterilization with Ethylene Oxide). Mean plasma volume processed by each IA procedure was 4.3 (2-5.5) times. No adverse events were observed during any IA procedure. All patients achieved successful desensitization with no Antibody-Mediated Rejection (AMR) reported. All patients continue to do well clinically with mean follow-up period of 6.5 (1-17) months. Although IA was expensive, it offered multiple advantages like specificity, larger plasma volume processing with faster reduction in titer, no ‘replacement fluid’ requirements and no adverse events in present case series.

Conclusion: IA plasmapheresis was universally successful in decreasing the ABO-isoagglutinin titers to desired level in all prospective SOT patients.

2019 May; 13(Suppl 1): S32–S100.

PP 24: Prevalence of transfusion reactions in a tertiary care hospital of north-east India

Background: Transfusion of blood and components can be a double edged sword which should be used judiciously. It can save lives; nevertheless it is not free from hazards. A transfusion reaction is defined as any transfusion-related adverse event that occurs during or after the transfusion of whole blood, blood components, or human-derived plasma products.

Aims: The present study aimed to find out the prevalence of acute transfusion reaction (TR) in patients of a tertiary care hospital of North-East India, who received blood and components in a two year period.

Methods: This prospective, cross sectional study was conducted in the department of Transfusion Medicine of RIMS, Imphal for a period of 2 years from September 2015 to August 2017. All the TRs were evaluated and classified in the blood bank using a preformed work up form and were notified to the ‘Haemovigilance programme of India’ (HvPI).

Results: A total of 30,936 blood component units were transfused during these 2 years, a total of 28 (0.09%) acute transfusion reactions were reported and evaluated as per departmental standard operating procedure, majority occurring in females and in 21-30 years age group. Among the types of TRs encountered, febrile non haemolytic transfusion reactions (FNHTRs) were most frequent (57.57%) followed by Allergic reactions (39.29%). Only one case each (3.57%) of sepsis and acute hemolytic reaction was encountered. TRs occurred most frequently with packed red blood cell transfusions.

Conclusion: The prevalence of TRs may underestimate the reality because of under reporting. With the help of Hospital Transfusion Committee, the study will facilitate the blood bankers and clinicians to formulate policies in the transfusion practice to make it better. Since the most common TR is FNHTR, processes like ‘universal leucoreduction’ of blood units can be implemented in the blood bank.

2019 May; 13(Suppl 1): S32–S100.

PP 25: A prospective interventional study to assess the impact of a “structured compact training” on knowledge and skills of safe blood transfusion practices among nurses working in a tertiary care institute

Background: Nursing officials have an important role in blood safety. It is highly essential to promote for improved quality and safety of transfusion practices in the nursing staff at the patient's bedside.

Aims and Objectives: To study the impact of training on administration of blood components after assessing the baseline knowledge in nursing staff.

Materials and Methods: In this prospective, interventional, single blinded study, participants were recruited from surgical wards-. Anonymity of the subjects was maintained throughout the study. After obtaining an informed consent, the nursing staff included in the study answered a validated questionnaire form which had 5 domains – A-Blood components, B- pre-transfusion checks, C- Transfusion process, D- post transfusion process and E-blood administration process. The nursing staff (intervention group) were given a training on the administration of blood components through a detailed lecture and a power point presentation. The influence of training on their knowledge, attitude and practices was evaluated through Kirkpatrick's four levels of training evaluation.

Results: Of the 84 subjects recruited, 54 completed the study. The mean age of the participants was 25.91±3.15 years (range 20-35 years). There were 30 females and 24 males. Of the 54 participants, 14 had less than one-year experience in service, 37 had 1-5 years’ experience, and 3 had more than 5 years’ experience. Before the intervention, the mean correct answers for domain A, B, C, D and E were 1.58±1.11 (range 0-4), 1.92±1.177 (range 0-5), 4.67±1.47 (range 2-8), 1.67±0.7 (range 0-3), and 3.87±1.92 respectively. Post the intervention, the scores improved significantly for domain A, B, C, D and E to 4.13±1.167 (range 0-5), 3.5±1.31 (range 1-7), 6.87± 1.48 (range 4-9), 2.7±0.47 (range 2-3), and 5.87±1.65 respectively.

Conclusions: Based on results of the study, we concluded that imparting information to the nursing staff significantly improved their knowledge about the transfusion services.

2019 May; 13(Suppl 1): S32–S100.

PP 26: Preoperative variables in overall blood component utilisation in liver transplantation

Background: Patients with end-stage chronic liver disease have a wide range of hemostatic abnormalities usually require blood transfusion during the procedure due to hemorrhage. As transfusion services remain an essential part of Liver transplantation, the ability to predict the preoperative variables will help blood transfusion services in improving preparedness and decrease wastage of limited resources.

Aim: To find out the preoperative factors that could predict the outcome of blood component utilization in end-stage chronic liver disease patients who had undergone liver transplantation.

Methods: The study was conducted in the Department of Transfusion Medicine, The Tamil Nadu Dr. M.G.R Medical University, Chennai for a period of 2 years by collecting retrospectively the available data of 20 patients who had undergone liver transplantation at Stanley Medical College Hospital, Chennai from April-2016 to March-2018. MELD score and preoperative factors like age, etiology, hemoglobin, platelet count were analyzed to find out the predictive correlation of these factors on blood component utilization.

Results: The total number of 68 units of PRBC, 55 units of FFP, 33 units of cryoprecipitate and 18 units of SDP were transfused intra-operatively during liver transplantation surgery.

The average numbers of PRBC, FFP, Cryoprecipitate and SDP units transfused respectively.

• MELD score 10-19 were 3.7,2.7,1.8&1, for the score 20-29 were 1.8, 1.9, 0.7 & 0.4

• Age ≤40 years 2,1.7,1&0.7, >40 years 4.1, 3.3, 2 & 1

• Etiology of ethanol related DCLD 4.7,3.2,2.1&1.1, Wilson Disease 2,1.6,1.2&0.6, Cryptogenic 3.2,3.5,1.8&0.7, HBV 2.5,2.5,1&1, HCC 2,2,1 & 1

• HGB ≤10 gm% 3.1,3,1.8&0.7, >10gm% 3.6,2.6,1.5 & 1

• Platelet count of ≤50,000/cmm 2.8,2.6,1&0.8, >50,000/cmm 3.6, 2.8, 1.8 &0.93.

Conclusion: In our study, the utilization of blood components during liver transplantation were more among patients above 40 years with ethanol related DCLD. It was observed that there was an inverse correlation between blood components utilization and MELD score/Platelet count. There were no significant difference between the blood component utilization and haemoglobin values above or below 10 gm%.

2019 May; 13(Suppl 1): S32–S100.

PP 27: Pregnancy loss and fibrinogen – A case report on role of cryoprecipitate

Background: Fibrinogen is a primary binding molecule, connecting platelets together via activated glycoprotein GPIIb/IIIa. Hypofibrinogenemia is an autosomal recessive inherited bleeding diathesis, it is associated with poor wound healing and recurrent miscarriage. Absence or significant decrease of fibrinogen in maternal serum is sufficient to cause rupture of maternal vessels and impaired trophoblastic infiltration causing bleeding and recurrent abortion. The available treatment uses cryoprecipitate, fresh frozen plasma and fibrinogen concentrate. The treatment during pregnancy is suitable only for patients correctly diagnosed with recurrent abortion.

Patient Concern: A case of 23 year old antenatal female married for 1 1/2 years with obstetric score of G2P1A1 with second trimester pregnancy loss and family history of recurrent abortions.

Diagnosis: Hypofibrinogenemia was detected after first pregnancy loss.

Outcome: During the entire pregnancy cryoprecipitate was transfused, twice a week 6 units of cryoprecipitate, the fibrinogen level was maintained above 70 mg/dl. Pregnancy was terminated at 36 weeks with emergency Cesarean Section in the view of maternal pyrexia, pre-operatively fibrinogen level was 97 mg/dl. Intraoperative 5 units of cryoprecipitate and Postoperative 4 units of cryoprecipitate were given, puerperal period was uneventful. Post-operative fibrinogen level was 139 mg/dl. She delivered a healthy male baby with no obvious bleeding tendencies.

2019 May; 13(Suppl 1): S32–S100.

PP 28: Application of autologous platelet rich plasma in treatment of a chronic nonhealing ulcer

Background: Chronic non healing ulcers are the major cause of non traumatic lower limb amputations. Application of platelet rich plasma is a cost efficient alternative to conventional treatment which increases rate of healing by providing growth factors released from alpha granules.

Aim: To evaluate the safety and efficacy of Autologous PRP for treatment of chronic non healing ulcer.

Case Discussion: 60 yr old female patient with history of diabetes mellitus type 2 presented to surgical OPD with an ulcer on right foot of 4 months duration. History of trauma to the right foot is present. Conventional treatment of wound debridement and dressing was done for 3 months but there was no improvement and planned for amputation. Informed consent was taken for topical application of Autologous PRP gel on the ulcer and is followed up. Autologous PRP is prepared from whole blood by density gradient centrifugation with fivefold increase in platelet count. Calcium chloride (1:9 ratio) and thrombin (500 IU/ml) are added to activate platelets and initiate release of growth factors. Within 5-10 seconds, a gelatinous material is formed which releases 70-80% of growth factors upon activation. Ulcer was debrided, cleaned with betadine solution and irrigated with normal saline; PRP gel is placed on the ulcer and covered with non absorbent dressing at bedside. Twice a week dressing is done for 5 weeks. Reduction of ulcer size (length*breadth), presence of infectious tissue, presence of pain and discharge, formation of granulation tissue, platelet count in PRP are the factors checked on every dressing. 100% resolution was seen at the end of 5th week.

Conclusion: This case report has demonstrated that Autologous PRP is a safe and effective way which enhances healing and prevents lower limb amputations. Further research and clinical trials on larger population are needed to validate the results.

2019 May; 13(Suppl 1): S32–S100.

PP 29: Maximum surgical blood ordering schedule for cancer surgeries - An institutional study

Over ordering blood is a common practice in surgeries. This can be corrected by a simple means of changing the blood ordering pattern. A retrospective study was carried out in the largest tertiary cancer care hospital of Gujarat for two years of period to study the blood ordering strategies in the hospital for surgical patients. The total units demanded and the corresponding units issued were studied for surgical patients. Thereafter, transfusion probability and ratio of units cross-matched to actual units transfused (C/T ratio) was calculated. Than transfusion guidelines for all surgeries requiring transfusions were proposed and implemented. Significant improvement was found in above three measures. The study also identifies the common cases where ‘Type and Screen’ (T&S) procedure could be introduced in cases where the transfusion probability is low. The implementation of this proposal will avoid over-ordering of blood which is beneficial to the institute.

2019 May; 13(Suppl 1): S32–S100.

PP 30: Prophylactic low dose platelets may be beneficial for the haematopoietic stem cell transplant patients

Prophylactic transfusions of platelet are the standard therapy to prevent bleeding in the patients with transient thrombocytopenia were being treated with high-dose chemotherapy followed by autologous haematopoietic stem cell transplantation (HSCT). Low to moderate dose of prophylactic platelet transfusions (<20,000/μl) not only prevent clinically relevant haemorrhage but also decrease in donor exposure and cost benefit due to single dose of SDP may be divided in two bags.

Aims: (1) Decrease multiple donor exposure related complication (2) cost effective.

Methods: As per our SOP of a single donor platelet (SDP) donor selection criteria, donor should be more than 60 kg weight, having good visible ante cubital vein, more than 150/cu mm platelet count, > 12.5 Haemoglobin, TTI screening negativity, not taken any anti platelet drugs and fulfil other criteria of blood donation. All the procedures done by the Optia machine. Time taken 60 to 70 minutes and achievable yields 3 -4 X 1011 which were divided in two doses.

Results: First Three (3) patients were MULTIPLE MYELOMA (2 male and 1 female) average age 54 years. Last two patients had refractory non Hodgkin's Lymphoma and Mantel cell lymphoma, and both were male, with their ages 21 years and 65 years respectively, cryopreserved HSC transfused after 1 month of peripheral autologous HSC collection. SDP platelets were approximately collected on the 5th and the10th day after HSC transfusion. Two bags were prepared from a single donor and average yields of platelets per bag were 1.5 to 2.2 X 1011 with volume 120 ml to 180 ml per bag. There was an average increment of platelets 40000/cu ml after single unit transfusion. None of the patients had any bleeding episode.

Remarks: Prophylactic low dose SDP may prevent bleeding in thrombocytopenic post SCT patients and it is cost effective.

2019 May; 13(Suppl 1): S32–S100.

PP 31: Role of a multidisciplinary team and massive transfusion protocol in improving patient outcome – A case study

Background: Management of a massive, life-threatening primary postpartum haemorrhage with DIC is a challenge for the clinical teams and hospital transfusion service as mortality is high and its etiology is multifactorial. The implementation of a standardized massive transfusion protocol with a multidisciplinary approach is necessary to prevent lethal triad of hypothermia acidosis, and coagulopathy.

Aims: The main aim of this case study is to highlight the importance of a multidisciplinary team approach and MTP protocol in improving patient outcome.

Case Study: The patient was brought to the emergency department of our hospital on 28th June in a very critical condition as a referred patient from another hospital. She had undergone a caesarean section for pregnancy with abruptio placentae with excessive bleeding on 27th June. Investigations showed her haemoglobin as 4.5 and a diagnosis of DIC (disseminated intravascular coagulation) was made. CT scan detected a massive haematoma inside her abdominal wall and a decision for surgery was taken. MTP was activated and 36 units of blood components 12 units PRBCs (packed red blood cells), 12 units of FFPs (fresh frozen plasmas) and 12 units of PCs (platelet concentrate) were transfused.

Results: Despite this massive blood transfusion, the patient recovered fully with overall 63 units of blood components transfusions and was discharged later with no complications. We followed standard blood transfusion protocol of our hospital with a ratio of 1:1:1 for PRBC, FFP and PC respectively. Each MTP pack included 4 units of PRBCs, 4 units of FFPs and 4units of platelets.

Conclusions: A damage control protocol should be prepared to manage patients who have significant acute blood loss. The rapid infusion of the correct ratio of blood products along with multidisciplinary team approach has been shown to improve patient survival and decrease overall usage of blood.

2019 May; 13(Suppl 1): S32–S100.

PP 32: Clinical determinants and pattern of red cell transfusion in a tertiary level neonatal unit

Background: Red cell transfusions constitute a crucial tool for the care of preterm infants. Shorter RBC life span, insufficient erythropoiesis and iatrogenic blood loss contribute to physiologic anemia of prematurity.

Objective: To identify the neonatal clinical factors that influence red cell transfusion pattern in preterm infants with anemia.

Methods: This retrospective study was conducted in neonatal ICU of SAT hospital, Trivandrum which is a tertiary care centre. Preterm neonates who received atleast one red cell transfusion were analysed. Data pertaining to antenatal and postnatal patient characteristics, indications for transfusions, number and volume transfused were obtained from the medical records.

Results: Out of 300 preterm neonates analysed 216 (72%) received atleast one PRC transfusion at the end of their hospital stay. Majority (57.4%) of the transfused were males (p value 0.126). Mean gestational age of transfused group was 29.7 weeks (nontransfused 32.1 weeks). and Mean birth weight was 1.29 +/- 0.47 kg (nontransfused-2.07 +/-0.39 kg) (p value-0.001). Mean haematocrit of the transfused group was 21. 4 +/- 4.2% and non-transfused group was 49.5 +/- 3.8%. Median number of RBC transfusions was 2 and mean volume of transfusion was 37.4 ml. Median postnatal day of transfusion was 15. Volume transfused was inversely related to birth weight and gestational age. Mean Iatrogenic blood loss was 9.7 +/-2.1 ml in the transfused group and 5.01 +/-1.3 ml in the non transfused. 50.5% and 45.4% of the transfusions were associated with administration of inotropes and significant mechanical ventilation, respectively. Prevalence of apnoea of prematurity was 43.5% in the transfused group vs 3.6% in nontransfused (p=0.0).

Conclusion: Clinical parameters like gestational age, birth weight, oxygen supplementation, pretransfusion haematocrit, administration of inotropes, iatrogenic blood loss, apnea of prematurity are significant variables helpful in identifying babies that are likely to require transfusions during the later phase of their anemia of prematurity.

2019 May; 13(Suppl 1): S32–S100.

PP 33: Rare phenotypes are huge challenge for blood banks: A case of Rh D-- phenotype

Red blood cells that do not express RHCE protein at their plasma membrane lack the C,c,E and e antigens are known as a Rare Rh Phenotype D--. We describe a rare Rh phenotype D-- in this case study.

Case Report: A 27 year female G4P3L1 admitted at 33 weeks with placental abruption with profuse bleeding PV and severe hypotension & tachycardia (Pallor +++ Pulse 136/min BP: 70/40 mmhg). Patient was managed by crystalloid infusion temporarily. Estimated blood loss around 1.5 liter. USG showed hydrops fetalis. She had lost two babies soon after delivery due to unknown cause. In emergency one O Negative unit was started for transfusion but had severe adverse reaction (hemolytic). Hb- 6.5 gm/dl Platelets- 74000 cells/mm3 WBC- 25400 Sr. Creat 1.9mg mg/dl Total Bilirubin 2.2 mg/dl (direct 1.08). Request of one Red cell unit was received at our blood bank. Blood group found O positive, antibody screen pan reactive, DAT Negative, autocontrol negative & cross match incompatible (+4) with all 15 O positive units. Antibody identification with 11 cells also pan reactive (+4). Rh Kell phenotyping found D+C-c-E-e-K- (Rh D--) by column agglutination. The tests were repeated by tube technique for confirmation & results were same. Being rare phenotype & multiple Rh antibodies patient didn’t get any compatible blood but managed without transfusion.

Conclusion: Rh D—is rare phenotype & transfusion for these patients is big challenge. There is need to develop Rare Donor Registry in our country to manage all Rare Phenotypes safely.

2019 May; 13(Suppl 1): S32–S100.

PP 34: A clinical audit of adverse reactions to blood transfusions reported to a stand-alone blood centre

Background: Establishing, monitoring & reporting protocols related to the process and outcome of transfusions can be very challenging to implement in a stand-alone blood centre. Our blood centre caters to more than 100 users for transfusion support.

Aim: To evaluate the compliance related to reporting of transfusion episode and further evaluate the frequency of reported transfusion reactions in period of 1 year and 8 months.

Materials and Methods: All transfusion reactions feedback forms received at blood centre during study period were evaluated for completeness of the information provided, type of transfusion reaction reported, type of blood component implicated, time of transfusion, location of blood transfusion in the hospital, pre & post transfusion monitoring and the staff who completed the form.

Results: Total 29,316 blood components were issued and 35 (0.12%) transfusion reactions were reported. 23 were male (65.71%) and 12 were female (34.28%), 3 were from paediatric age group (8.57%) & 32 (91.42%) were adults. 6 (17.14%) transfusions were done in night. 32 (91.42%) of transfusions were given in the in-patient sitting. 18 (51.42%) transfusion reactions were mild, 7 (20%) were moderate and 6 (17.14%) were severe. 22 (62.85%) transfusion reactions were associated with PRBC, 4 (11.42%) with platelets and 8 (22.85%) with plasma transfusion. Pre & post Transfusion assessment was done in 34 (97.14%) cases. Febrile non-hemolytic transfusion reactions were the commonest (50%) reactions reported. Incomplete information was found in 7 (20%) cases.

Conclusion: FNHTR was the commonest transfusion reaction reported and severe transfusion reactions were uncommon. Transfusion reactions were more associated with PRBC. Universal leucodepletion may result in significant reduction of transfusion reactions in our recipients. Education of personnel who fill up the transfusion reaction forms is important for better post-transfusion reaction work-up.

2019 May; 13(Suppl 1): S32–S100.

PP 35: Effect of red blood cell storage age on posttransfusion haemoglobin increment

Background: Biochemical and structural changes that occurs in RBCs during the storage may have a potential clinical relevance, which is still unclear. Also, there has been data that 15-25% of the old red cells are removed from the circulation within 24 hours after transfusion. So this study attempts to find the effect of storage age on efficacy of PRBC transfusion.

Aim: To study the relation between RBC storage age and post transfusion Hemoglobin increment in recipient.

Methods: In this prospective study, data were obtained from consecutive patients admitted in the department of medicine who received single packed red cell transfusion in 24 hours period and were followed up for post transfusion hemoglobin. Patients with evidence of hemolysis, active bleeding and receiving concurrent IV fluid were excluded. Post transfusion hemoglobin increment was compared between fresh [Category I, <7 days] and older [category II, >7 days] PRBC units.

Results: Data of 100 patients corresponding to 100 PRBC units were analyzed. In category I, Mean age of recipients was 58.27 years, mean pre-transfusion Hemoglobin was 5.17 g%, mean post-transfusion Hemoglobin was 6.75 g%. In category II, it was 57.10, 5.15 and 6.38 respectively. There was a negative correlation between storage age [M=4.21, SD= ±4.15] and Hemoglobin increment [M= 1.51, SD=±0.54] with r = .380, p = .000. Negative correlation between Pre-transfusion Hemoglobin and Hemoglobin increment [r= .398, p= .000] was also observed. No correlation found between age of donor and Hb increment in recipient. Both categories II and I showed an adequate increment [1.23 g%, 1.58 g% respectively] but there was a reduction in post-transfusion Hemoglobin increment in the category II [t=2.75, p= .007].

Conclusion: Post transfusion Hemoglobin increment decreases with increase in storage age of PRBC units. Yet an adequate increment is obtained with transfusion of both fresher and older RBCs.

2019 May; 13(Suppl 1): S32–S100.

PP 36: Intrauterine red cell transfusions - A look back

Introduction: Intra-uterine transfusion (IUT) with red cells: Main-stay of treatment for fetal anemia.

Aim: Retrospective analysis of red cell IUT over the period Jan 2014-June 2018.

Materials and Methods: This is a retrospective study of 53 patients who received red cells for IUT at our centre. Leukoreduced, irradiated O Negative Red Cell Concentrates (RCC), antigen negative for the corresponding maternal antibody with a high Haematocrit (HCT) of over 80% (mean 83.4%, range 75-91%), were transfused. Sickling test was performed on the units used. Volumes transfused varied depending on gestational age and fetal weight.

Results: Of the 53 patients 34 (64%) received IUT for fetal anemia due to immune causes. Anti-D alone was implicated in 17cases, dual antibody D &C in 16 cases and anti-D, anti-C with anti-Jka was detected in 1 patient. Anti-D titres ranged from 16 to 2048 while anti-C titre ranged from1 to 16. Number of IUT's ranged from 1 to 4 per patient with interval ranging from 7 days to a month between transfusions. Antibody screening was negative in 7 patients (15 %) and could not be worked–up in the remaining.

Discussion: This study reinforces that fetal anemia caused by anti-D continues to be the major indication for IUT, necessitating the need for better Rh (D) immunoprophylaxis. Further, significant fetal anemia was noted in 5 cases where anti-D titre was <32 indicating need for correlation with fetal Middle-cerebal artery Peak systolic velocity (MCA-PSV) blood flow. Non-immune causes for IUT were G6PD deficiency and Parvo-virus infection. Other causes for IUT to be looked for are, red cell membrane defects, red cell enzyme defects and Haemoglobinopathies.

Conclusion: Red cell antibody screening in pregnancy, adequate immunoprophylaxis with anti-D followed by provision of antigen negative blood for IUT will go a long way in the management of Haemolytic disease of Newborn.

2019 May; 13(Suppl 1): S32–S100.

PP 37: Sometimes the best way forward is reverse - A case report

Background: ABO discrepancies are not uncommon to Immunohaematologists. Any discrepancy should be carefully evaluated alongside a detailed clinical history as aetiology implicated in a discrepancy could aid the clinician. Here we describe a case where pursuing of clinical details of a patient with ABO discrepancy led to a possible clinical diagnosis.

Case Report: A request was received for a unit of red cells at our centre for a 19-years old female with anaemia, being planned for bronchoscopy. The blood grouping results showed the forward typing to be suggestive of O Rh positive. However, reverse grouping showed completely negative reactivity with A, B and O cells. Reverse grouping was repeated with methods to enhance the sensitivity including variable temperatures and incubation times on both the tube and CAT technology but yielded identical results. The complete absence of isohaemagglutinins raised the possibility of an immunodeficiency syndrome. A detailed history was subsequently obtained which revealed recurrent pulmonary and gastrointestinal infections since childhood. With this combination of clinical and laboratory features, Ig levels were suggested and subsequently done, which revealed features consistent with Common Variable Immune Deficiency (All Ig isotypes <10mg/dL, no T-cell deficiency and absence of secondary hypogammaglobulinemia).

Conclusion: Combining clinical features with laboratory findings is critical to optimal patient care. While documenting discrepancies in ABO grouping, it is critical to interface with clinicians to assess if there is any primary clinical aetiology which is driving the discrepancy identified. In this case, we highlight the role of reverse grouping in guiding further investigations which led to a definitive diagnosis and thus appropriate intervention.

2019 May; 13(Suppl 1): S32–S100.

PP 38: Transfusion practice in acute promyelocytic leukemia patients – Experience from tertiary care cancer center

Introduction: Acute promyelocytic leukemia (APML) is a myeloid leukemia subtype associated with a high mortality rate in newly diagnosed patients. Transfusion support is integral to patient care and substantial amounts of blood may be required per patient. Understanding the transfusion requirements of these patients is crucial for planning health care strategies, including blood component provision and donor recruitment.

Methodology: Retrospective study of six APML patients who underwent induction chemotherapy was analyzed. Patients were stratified into low, intermediate and high risk categories on the basis of WBC and platelet count. Laboratory values and blood transfusion requirements till their hospital stay were collected.

Results: Five out of six patients presented with bleeding and coagulopathy at presentation. Liberal transfusion thresholds were followed for all three categories. The nadir platelet count for platelet transfusion was 16,000±4335cells/μl. We observed no significant differences in blood usage between the groups. The average blood consumption was PRBC (5.5±3.7 units), RDP (3.1±3.1 adult dose); SDP (6.3±5.0 adult dose), FFP (37±32 units), Cryo (51±49 units). The mean interval between the transfusions for PRBC was 5.5±2.1 days and for Platelets 2.2±0.1 days. The average length of stay for these patients was 42±25 days.

Expected Conclusion: We observed the most important predictor for transfusion may be disease burden and not risk stratification. By increased use of apheresis platelets, number of donors required for platelet support was minimized. Accurate assessment of transfusion requirements of this group will help in planning therapy and directing resources for new patients.

2019 May; 13(Suppl 1): S32–S100.

PP 39: A survey for assessing the knowledge of transfusion medicine among resident doctors, interns and staff nurses in a tertiary care centre, Kerala

Background: The knowledge of blood transfusion is essential for safe transfusion practices. Previously a few studies had investigated this topic which showed deficiencies in both knowledge and practices among the interns, resident doctors and nurses.

Aims and Objectives: To assess the knowledge and practices in blood transfusion among resident doctors, interns and nurses in JMMC & RI.

Methodology: A descriptive cross sectional study using a self administrated questionnaire comprising of 45 questions from transfusion medicine. A sample size of 150 participants is being considered. Questions are divided into 5 sections which includes : Sec A- Awareness about blood donation, Sec B: Blood donation, Sec C- Components and blood processing, Sec D- Storage of blood, Sec E- Blood Transfusion practices.

Results: Since the study is going on results cannot be produced now and will be sent later.

2019 May; 13(Suppl 1): S32–S100.

PP 40: Incidence of hyperfibrinolysis in liver transplantation and its reversal by tranexamic acid

Liver transplantation has been accepted as an effective treatment for patients suffering from end stage liver disease due to various etiologies. Major blood losses is a known complication in liver resection and liver transplantation. Hyper-fibrinolysis plays a significant role in blood loss requiring massive transfusion during anhepatic phase. The use of Tranexamic acid (TXA) prevents traumatic and perioperative bleeding by its antifibrinolytic action. However, a large dose of TXA may increase the risk of thrombosis.

Aim: To find out the incidence of of hyperfibrinolysis in liver transplantation and its reversal by Tranexamic acid To find out the incidence of small vessel thrombosis associated with Tranexamic acid treatment in liver transplantation patients.

Materials and Methods: The study was conducted in the Department of Transfusion Medicine, The T.N. Dr. M.G.R Medical University, Guindy, Chennai for a period of 2 years by collecting retrospectively the available data of 20 patients who had undergone liver transplantation at stnley medical college, Chennai. The factors included in the study were the etiology of End stage liver disease, the effect of Tranexemic acid on reversal of Hyperfibrinolysis and incidence of small vessel thrombosis.

Results: The incidence of Hyperfibrinolysis was observed in 16 cases (80%) by thromboelastogram. The tranexemic acid used for all the 16 patients showed complete reversal by TEG. Two of these patients treated by tranexemic acid developed portal vein thrombosis, however with effective intervention the patients survived. The primary etiology for these 2 cases respectively was etahalol related DCLD and Wilson disease.

Conclusion: In our study, we observed the use of tranexamic acid in hyperfibrinolysis is very effective. However, timely recognition and effective intervention is always necessary to manage small vessel thrombosis, one of the dreaded anticipated complications of its use.

2019 May; 13(Suppl 1): S32–S100.

PP 41: Characterisation of venom induced consumption coagulopathy in patients with haemotoxic snake bite and the effects of blood products on coagulation parameters

Introduction: Snakebite is one of the most important “Neglected Tropical Diseases” in terms of both incidence and severity, and its clinical characteristics. Venom-induced consumption coagulopathy (VICC) is the core pathogenic mechanism in haemotoxic snakebites. The common derangements seen are prolonged Prothrombin time (PT), prolonged Activated Partial Thromboplastin Time (APTT), low/undetectable fibrinogen. VICC is characterized by reduction of coagulation factors and the absence of systemic microthrombi and end-organ damage. The time course in VICC is rapid- occurring within a few hours of envenomation and resolution within 24-48 hours, if treated appropriately.

Aim: This study focuses on characterizing the effects of snake bite on the haemostatic system, transfusion requirements and effect of transfusion on haemostatic parameters

Materials and Methods: This was a retrospective study conducted between January 2012 to December 2016 at Christian Medical College, Vellore.

Results: Data from 576 potential snake bite victims were analysed, out of which 280 (48.6%) patients had a haemotoxic snake bite. Among these 47 (16.8%) patients were transfused blood and plasma products. There was a male preponderance among patients (70.2%). Isolated haemotoxic features and local reaction were seen in 8.5%. Coexisting neurological and/or renal manifestations were seen in more than 90% patients. The average dose of ASV received per patient was 18.9 ±7.75 vials. Baseline INR more than 1.5 was seen in 87.2% and an elevated APTT ratio was seen in 55.3%. All components were transfused: platelet concentrates (30%), FFP (66%), cryoprecipitate (32%) and cryosupernatant (11%). Mortality was 10.6% among this patient group with transfusions.

Discussion and Conclusion: Snakebite was and still remains a problem that can easily be tackled, if diagnosis and treatment is given in a timely manner. The role of a good haemostasis laboratory in detecting VICC and management of the patient is emphasized by this study.

2019 May; 13(Suppl 1): S32–S100.

PP 42: To evaluate the role of therapeutic phlebotomy in decreasing hemoglobin and preventing complications/relieving symptoms in primary and secondary polycythemia patients

Background: Therapeutic phlebotomy is the preferred treatment for blood disorders in which the removal of RBCs or serum iron is the most efficient method for managing the symptoms and complications. It is currently indicated for treatment of primary and secondary polycythemia, hemochromatosis, sickle cell diseas, etc.

Aim: The main aim is to evaluate the role of therapeutic phlebotomy in decreasing Hb and preventing complications/relieving symptoms in primary and secondary polycythemia patients.

Methods: A retrospective analysis of patients who underwent therapeutic phlebotomy from January to August 2018 was done. The data regarding patients diagnosis, desired Hb, amount of blood withdrawn and decrease in Hb, whether replacement fluid given or not were obtained from patient's records.

Results: A total of 15 patients underwent therapeutic phlebotomy during this study period. Out of which 7 patients (46.66%) had primary polycythemia due to myeloproliferative disorder with Jak 2 mutation positive in 6 patients and negative in 1 patient. Remaining 8 patients (53.33%) had secondary polycythemia due to chronic obstructive lung disease, cyanotic heart disease, chronic kidney disease. 4 patients with primary polycythemia underwent the procedure 4 or more times and 3 patients underwent less than 2 times. Following the procedure, Hb level decreased and thrombotic complications were not reported in any of these patients. Whereas out of 8 patients with secondary polycythemia, 6 patients underwent the procedure only once and patients relieved of symptoms post procedure along with decrease in Hb level.

Conclusion: Our data supports that the therapeutic phlebotomy plays a major role in treatment of primary polycythemia than in secondary polycythemia.

2019 May; 13(Suppl 1): S32–S100.

PP 43: Role of therapeutic plasma exchange in various clinical conditions and evaluating the outcome - A single center study

Background: Therapeutic Plasma exchange (TPE) removes the offending substance like autoantibodies from plasma and has shown significant benefits in autoimmune conditions like Gullian Barre syndrome, Myasthenia gravis, Thrombotic thrombocytopenic purpura etc.

Aim: To evaluate the efficacy of Therapeutic plasma exchange (TPE) in various clinical conditions.

Methods: Prospective Observational Study. Patients with various clinical condition who required and underwent TPE for various indications from June 2014 to July 2018 were analyzed for efficacy of TPE.

Results: 42 patients were studied, of which 10 patients were diagnosed with GBS, 9 patients showed significant improvement in muscle power, mobilized and discharged after 5 cycles of TPE, 1 underwent more than 10 cycles and showed minimal improvement. Out of 9 patients with myasthenic crisis, 7 patients improved after mean 4 cycles of TPE, 2 patients died of respiratory failure. Of 6 patients with rapidly progressive glomerulonephritis who were on TPE along with dialysis, 4 patients showed significant improvement and 2 didn’t improve. Of 5 cases of antibody mediated rejection- post renal transplant, 4 showed improvement in renal function and 1 went into complete rejection. Out of 4 cases of CIDP, 3 showed improvements and 1 died. Out of 3 patients with severe sepsis 2 improved after 4 cycles of TPE and 1 died. 2 patients with Hemolytic uremic syndrome who underwent TPE showed significant improvement. One patient with lupus nephritis underwent TPE and without any significant improvement. 1 Patient with Neuromyelitis optica and 1 patient with Wegener's granulomatosus improved signicantly after 4 cycles of TPE.

Conclusion: Therapeutic plasma exchange can be applied in variety of clinical conditions and can be used as primary or second line therapy. Correct patients should be identified and timely TPE can bring definite improvement in patient clinical condition and can reduce mortality significantly.

2019 May; 13(Suppl 1): S32–S100.

PP 44: Whole blood exchange-an effective treatment modality for naphthalene ball ingestion induced methemoglobinemia: A case report

Background and Objectives: Naphthalene is a widely used chemical in domestic and commercial applications in the form of mothballs. However, intentional mothball poisoning has been rarely reported in the medical literature despite its widespread use in Indian households. Naphthalene poisoning results in haemolytic anemia and methemoglobinemia, intravascular hemolysis being marked particularly in those with G6PD deficiency who have low tolerance to oxidative stress.

Methods: A 21 year old male with alleged history of 1 mothball intake with suicidal intention 2 days back was referred to our department for whole blood exchange transfusion from department of Internal Medicine of our institute. His chief complaints were pain abdomen and reddish-black urine for 2 days, Jaundice and shortness of breath for 1 day. Investigations showed a low Hb of 8.3g/dL, markedly raised level of MetHb:19.6% and Total Serum Bilirubin: 19.8 mg/dL. Whole Blood Exchange Transfusion (WBET) was performed with Cobe Spectra® (TerumoBCT, LakewoodColorado, USA) by using TPE kit. Plasma was collected in plasma bag while RBCs were collected in a wastebag attached to return line of the disposable kit for discard. A separate peripheral return line was secured for transfusion of reconstituted whole blood units. A total of 6 pRBCs and 12 FFP units were used to replace one blood volume of the patient.

Results: After a single procedure of WBET, MetHb levels dropped from 19.6% to 4.6%, SpO2 rose from 66% to 90%. Patient became symptomatically better. No adverse event related to procedure was observed. He was given supportive management with IV Ascorbic acid and N-acetyl cysteine and advised hemodialysis in view of acute kidney injury (B.Urea:183 mg/dl, S.Creatinine:5.2 mg/dl). Patient was discharged following clinical improvement on Day 10 with advice for follow up at 3 months for G6PD levels.

Conclusion: WBET is an effective and safe treatment modality in patients with acquired methemoglobinemia secondary to mothball ingestion.

2019 May; 13(Suppl 1): S32–S100.

PP 45: Transfusion reaction or not? An unusual presentation

Background: Anaphylactic reactions in the context of transfusion is an extremely rare phenomenon. However, it's most characteristic presentation is an extremely severe reaction with the transfusion of a few ml of the implicated unit. Severe reactions, when they occur outside the previously observed and defined grids pose a challenge in assigning etiology to that reaction. We describe here such a challenging scenario.

Case Report: A 57-year old male admitted under ENT department was posted for nasal surgery. A request for two units of blood for the procedure was received by the blood bank. There was no history of prior transfusion nor any atopic symptoms. The patient received two units of whole blood and after the transfusion of 350ml of the second unit, the patient developed an anaphylactic reaction. The patient required reintubation and was managed aggressively with antihistamines, steroids and ionotropes and recovered subsequently. As a follow-upm, serum IgE levels, IgA levels and tryptase levels were done. IgA levels in the pre-transfusion sample was found to be slightly reduced (108.2 mg%) [Normal: 140-420 mg%] and IgE and serum tryptase levels in the post-transfusion sample were elevated.

Discussion and Conclusion: What is clear is that the patient had an anaphylactic reaction. However, the fact that the patient tolerated more than 1 unit of blood without clinical events, again raises the question about it being related to transfusion. Given the documented levels of IgA, it is unlikely to be related to an IgA deficiency though ideally levels of IgA subclass and anti IgA antibodies should have been measured. It is critical that patient be evaluated for other etiologies that could trigger an anaphylactic reaction. However, it is advisable that if repeat transfusion is required, that it be limited to washed cellular products with extremely close clinical monitoring.

2019 May; 13(Suppl 1): S32–S100.

PP 46: A study on assessing relationship between neonatal gestational age and packed red blood cell requirements

Introduction: Neonatal physiology varies with the maturity, age, weight and the presence of morbidities, it is difficult to formulate one parameter to guide all transfusion decisions. The increased risk of transfusion transmitted infections including unidentified infectious agents is a major concern in multiply transfused neonates. Moreover, neonates are the longest survivor of blood transfusion and can manifest adverse events with multiple donor exposure. So, the present study was conducted to assess the relationship between Gestational Age (GA) with Packed Red Blood Cells (PRBC) requirements along with number of donor exposure due to PRBC transfusions.

Materials and Methods: Retrospective study conducted for one year (September 2016 - August 2017). To assess the relationship between GA and PRBC transfusion, GA was categorised into 4 groups according to WHO & ACOG criteria viz Extreme preterm (<28 weeks), Very preterm (28 to <32 weeks), Preterm (32 to <37 weeks) and Term (>37 weeks). Number of donor exposure for each neonate due to PRBC was also noted.

Results: 66 neonates received 129 packed red cell units. The mean (±SD) PRBC requirement of 4 groups were Extreme preterm (2.16 ± 1.47), Very preterm (2.45 ± 1.43), Preterm (1.65 ± 1.52) and Term (0.80 ± 0.97), p= 0.00. PRBC requirement was inversely proportional to the gestational age of the neonate. The mean donor exposure was 1.29 per neonate (range 1-5).

Conclusion: Very low birth weight premature neonates bear the brunt of multiple donor exposures due to frequent PRBC transfusions. Preterm infants of very low birth weight have very little iron stores and a small circulating volume of RBCs when they exit the uterine stage. Recently, BCSH illustrated an example algorithm for pedipack allocation for neonates according to gestational age for reducing donor exposure.

2019 May; 13(Suppl 1): S32–S100.

PP 47: An audit on single-unit transfusions and hemoglobin trigger

Introduction: The NICE guidelines for blood transfusion and the PBM recommendations state that a single PRBC unit should be the standard dose for patients with stable anaemia who are nonbleeding. The BCSH guidelines recommends that PRBC transfusion should be used to maintain Hb level of 7–9g/dl which is supported by AABB guidelines which recommends against transfusion for stable non-bleeding patients with Hb >7–8g/dl. Studies have shown that changing clinical transfusion practice can be difficult.

Aim: To audit the single unit transfusions in non-bleeding stable patients admitted in the Department of General Medicine.

Methodology: The study is conducted in the Department of IHBT and General Medicine in a medical college over a period of 2 months. Data collection includes All PRBC transfusions given to general medical in patients. A single unit transfusion definition was single PRBC unit transfused followed by a measurement of a post-transfusion Hb and patient assessment.

Results: A total of 2012 patients were admitted during the period. Only 204 of them required PRBC transfusion of which 43.13% females and 56.86% males. The median age was 55 years (17–89). A total of 280 PRBC units (1.37U/patient) were transfused.164 patients were given single unit PRBC transfusions. The most common indication was anemia. Length of stay when compared with those receiving multiple transfusion was less in the ones receiving single unit. The mean Hb trigger for those receiving single unit transfusions was 7.36 g/dl (3-8.8). Further evaluation is under going.

Conclusion: This study shows that transfusion rates are already lower compared to othere studies, which in turn reduces the potential risk to patients from allogeneic transfusion and cost/time associated with transfusion by adopting the key red cell recommendations from 2015 NICE Guidelines in blood transfusion. Further health economics analysis on the implementation of this single unit transfusion policy is being completed.

2019 May; 13(Suppl 1): S32–S100.

PP 48: A case of autonomic autoimmune ganglionopathy treated with therapeutic plasma exchange

Background: Autoimmune autonomic ganglionopathy (AAG) is a rare, acquired, immunoglobulin mediated disorder of autonomic failure due to autoantibodies to the nicotinic acetylcholine receptor of the autonomic ganglia (nAChR). The clinical picture manifests as pandysautonomia including orthostatic hypotension, recurrent syncope, anhidrosis, sicca syndrome (xerostomia and xerophthalmia), bowel and bladder hypomotility, and papillary dysfunction, although all manifestations are not present in all patients. We present a case report of a 40 year old male patient who presented with features of AAG and was successfully treated using plasma exchange (PLEX) followed by immunosuppressive therapy.

Aim: To report response to therapeutic plasma exchange in a patient with Autoimmune autonomic ganglionopathy.

Methods: Patient presented with features of autonomic dysfunction since one year Autonomic testing revealed severe sympathetic as well as cholinergic dysfunction. Patient was managed with five alternate day cycles of therapeutic plasma exchange under strict clinical monitoring.

Results: Patient tolerated the procedure well. Patient's orthostatic symptoms improved along with bladder, bowel and other symptoms. Following plasma exchange patient was started on steroids. Compression stockings and orthostatic precaution was advised.

Summary: The optimal therapy for AAG remains uncertain. No randomized controlled trials are available (to the best of our knowledge), and there are only limited case reports of successful treatment of AAG. Standard treatments for orthostatic hypotension including volume expansion, vasoconstrictors, compression stockings and abdominal binders, rarely provide adequate symptomatic relief in AAG. PLEX followed by immunosuppressive therapy resulted in sustained clinical improvement in this patient.

2019 May; 13(Suppl 1): S32–S100.

PP 49: Role of therapeutic plasma exchange in multiple myeloma patients with cast nephropathy

Background: Cast nephropathy develops in 30-80% of patients with renal disease due to multiple myeloma. Progressive obstruction in distal renal tubules accounts for irreversible decline in renal function. Therapeutic plasma exchange (TPE) has supportive role in oliguric patients who excretes >10 gm light chains in a day and whose creatinine become >6 mg/dl. Recently guideline in American society for apheresis 2016 had mentioned TPE as category III indication for patient with cast nephropathy due to multiple myeloma.

Aim: To assess efficacy of TPE therapy in patients having cast nephropathy.

Methods: A retrospective analysis of all TPE procedures was done over a period of 10 years (2006-2016). Procedures were done on different apheretic devices (CS-3000 Plus, Baxter, USA and Cobe spectra, Terumo BCT, Lakewood Co.USA). Patients’ pre and post procedural renal function parameters were analyzed by applying paired T test to assess the efficacy of TPE.

Results: A total of 11 patients with myeloma kidney having cast nephropathy were undergone therapeutic plasma exchange procedure with the aim to remove excess free light chains. The mean age of these patient was 51.54 year with a range of 33 to 77 years in a male to female ratio of 8:3. Rise in creatinine was presenting complaint in all these patients despite on chemotherapy. There was significant decline (p value of 0.043) in mean serum urea concentration post procedure from 133.45±49.59 to 99±45.07 mg/dl. Similarly significant decline in mean serum creatinine was observed post TPE from 4.85±2.83 mg/dl to 3.71±2.12 mg/dl with a p value of 0.013.

Conclusion: This retrospective study suggests that TPE has its supportive role in preventing irreversible renal dysfunction in patient with multiple myeloma.

2019 May; 13(Suppl 1): S32–S100.

PP 50: Cryopreservation of peripheral blood stem cells at a tertiary care hospital

Background: Peripheral Blood Stem Cell (PBSC) transplant is used for treatment of many malignant and non-malignant conditions. Cryopreservation is the technique used to preserve stem cells viable for longer duration, if the transplant/transfer of stem cells from donor to recipient is not done within 72 hrs. Dimethylsulfoxide (DMSO) and Hydroxyethylstarch (HES) are the most commonly used cryoprotectants. Cryopreservation is done either by using liquid nitrogen or controlled-rate mechanical freezers. This is proven safe and not associated with significant adverse effects like failure to engraft or GVHD.

Aim: To enumerate the role of Transfusion medicine in cryopreservation of stem cells.

Methods: This study has been carried out in the Department of Transfusion Medicine from April 2016 to April 2018 at Sri Ramachandra Medical College & Research Institute, Chennai.

Results: In the 2 years period from April 2016 to April 2018, total of 19 (Autologous: 11, Allogenic: 6 & Matched unrelated donor (MUD):2) PBSC were harvested among which for 6 cases, cryopreservation was performed. Among the 6 cryopreserved indications were (Autologous: 1-Neuroblastoma), (Allogeneic [3] Beta-Thalassemia 2 cases & SCID 1 case), (MUD: 2 – AML, Infantile ALL). DMSO (Dimethyl Sulfoxide) was used as cryopreservative agent, and stem cells are mixed with cryoprotective agent in 1:1 ratio. After processing, cryopreserved stem cells were stored at -800C in mechanical freezer. Just before infusion the cryopreserved stem cells were thawed at 370 C in a water bath.

Conclusion: Cryopreservation is the use of very low temperatures to preserve structurally intact living cells and tissues. In our study all patients received myeloablative chemotherapy before PBSC transplant. All patients demonstrated early engraftment and good recovery rates. Preservation of PBSC in mechanical freezers at <-800C is an easy and robust cryopreservation method which allows for a viable storage period of upto 3 months.

2019 May; 13(Suppl 1): S32–S100.

PP 51: A significant improvement in autonomic sensory and motor function due to stem cell application in old spinal injury

Background: A 21 years female post graduate student had a fall from a three storeyed building three years before with unconsciousness. She was admitted immediately in local hospital. Her vertebral X ray revealed multiple fracture of D11 to L2. She was operated on 4th day of injury for decompression and internal fixation and released after 3month of hospital stay. Her paraplegia, bilateral muscles wasting of lower limb with loss of voluntary control urinary bladder were persisting throughout. Her sensory, motor and cognitive function were satisfactory above the umbilical region. She was evaluated on Feb 2018 at our Institute of Regenerative Medicine and advised for autologous stem cell therapy injected subcutaneously on the site of injury after counselling and consent.

Aim: Autologus stem cell and platelet rich plasma (PRP) injection at the site of injury may help motor, sensory, cognitive function below the site of injury.

Methods: Autologous 15 ml of bone marrow aspirated after local anaesthesia by 2% lidocaine from posterior iliac crest in aseptic condition in a sterile EDTA containing vacutainer at 24°C. The Vacutainer centrifuge at 3000 rpm for 5 minutes to collect buffy coat settlement. An autologous PRP was prepared from 10 ml of blood collected from antecubital vein in aseptic condition and 5 ml of suspension (3 ml buffy coat and 2 ml PRP) prepared. The suspension was drawn in 10 ml syringe and inject subcutaneously at 4 weeks interval in 5 times along with advised for regular physiotherapy, using ankle splint and walker for standing with support.

Results: No obvious visible changes of improvement noted until 4th procedure. After regular physiotherapies and autologous buffy coat with PRP injection, it was noted that she gained some muscle power in the lower limb and was able to stand up with the support of walker after 4th cycle.

2019 May; 13(Suppl 1): S32–S100.

PP 52: Evaluation of reasons affecting the factor viii levels in hemophilia patients after therapy

Background: Hemophilia is an X-linked congenital bleeding disorder caused by a deficiency of coagulation Factor VIII (FVIII) (in hemophilia A) or factor IX (in hemophilia B). Replacement of a deficient clotting factor is a basic treatment. Inhibitor development is a significant treatment complication which decreases the effectiveness of replacement therapy and affects the quality of life.

Aims: Aims of this study were to evaluate the reasons affecting the FVIII level in hemophilia patients after treatment. Prevalence of FVIII inhibitor and effects of type of clotting factor used for treatment, severity of the disease and other causes in development of inhibitor were also evaluated.

Methods: With prior consent and history, blood samples collected from hemophilia patients were tested for PT, APTT, FVIII activity and FVIII inhibitor screening. FVIII inhibitor assay was performed on samples with positive FVIII inhibitor screening. All plasma based coagulation tests were performed using sodium citrate anticoagulated platelet poor plasma on automated coagulation analyzer with commercially supplied reagents. The test results and clinical history were statistically evaluated.

Results: FVIII inhibitor was developed in 24 patients (16.78%) out of 143 patients in this study. Out of 65 patients diagnosed of hemophilia A before 6 months of age, 18 patients (p value-0.001) had developed inhibitor against FVIII. Out of 79 patients taking “on demand” treatment, 18 patients (p value-0.032) developed inhibitor against FVIII. Out of 111 patients with severe hemophilia A, 24 patients (p value-0.003) developed inhibitor against FVIII.

Conclusion: Development of FVIII inhibitor is the most important factor affecting treatment of hemophiliaA patients. Patients with severe hemophilia, exposed to FVIII concentrates during first 6 months of life and patients taking “on demand” treatment are at more risk for inhibitor formation. Type of FVIII (cryopreciptate/recombinant FVIII concentrates) used for treatment has no significant effect on inhibitor development.

2019 May; 13(Suppl 1): S32–S100.

PP 53: Blood banking to bone banking: Eight years experience of gamma irradiated bone allografts

Background: Tissue banks serve as procurement and distribution centers of human tissues. These are provided as non-viable allografts, preserved by freeze-drying and sterilized by gamma irradiation. Many Blood Centres across the world provide services not only related to blood & components but also various tissue allografts. At our blood centre we started Tissue Bank in 2010 & process various bone allografts.

Methods: Bone tissues are collected after knee & hip replacement surgeries with the consent of patient & blood specimen with cold chain. The donor is evaluated and blood sample is tested for HIV, HBsAg, HCV & syphilis. Three levels of processing: 1. Wet 2. Dry 3. Termial Gamma Irradiation. Wet processing includes heat & chemical treatment including pasteurization at 60C for 3 hours & ethyl alcohol treatment. Dry processing includes lyophillization at -52C for 72 hour and Terminal Sterilization is done by Industrial Gamma Irradiation for the dose of 25000 Gy. The quality control tests are Bio Burden & moisture content test.

Results: From June 2010 to June 2018, we have collected 3481 bone allografts from various hospitals in & around Bangalore. Total 252 samples were rejected due to sero positivity, 31 rejected due to other reasons. We have issued 3022 freeze dried & 64 frozen bone allografts. Out of 3022, femoral heads were 874 &tibial slices were 2148. We have not received any adverse incidence due to allograft implantation.

Conclusion: The Gamma Irradiated Bone allografts effective, facilitating the formation of new bone and used for joint revision surgery & various reconstruction surgeries. The availability of safe, clinically useful and cost effective grafts have resulted in changes in surgical treatment. Like developed countries Blood Centres in our country can fulfil the increasing demand of tissues by aquiring adequate infrastructure & technical expertise.

2019 May; 13(Suppl 1): S32–S100.

PP 54: In-house cryopreservation of peripheral blood stem cell: initial experience from a tertiary care hospital

Background: Cell therapies based on hematopoietic stem cells (HSCs) have become the standard of care for a large number of clinical indications, and the number of patients and disorders being treated using HSCs continue to grow. Here, we report an analysis of first 10 cryopreservation of peripheral-blood stem-cell (PBSC) at our center in terms of CD 34+ viability and cell engraftments in those patients.

Methods: Peripheral blood stem cells (PBSCs) were collected from auotologus donors using P1YA kits on Com.tec (Fresenius kabi). A solution made up of 10% DMSO, 20% human serum albumin (HSA) with 6% HES was used for cryopreservation of cells. Whole procedure was done under fully aseptic technique using laminar air flow. A mechanical freezer (Thermo Scientific) was used to store stem cells at -800 Celsius.

Results: All 10 patients were posted for re-transplant (2nd/3rd) and heavily pre treated with chemotherapy. Average time duration of storage for cryopreserved stem cells was 27 days (range 10 to 56 days). The initial yield of the products before cryopreservation ranged from 3 X 106 to 10.2 X 106 per Kg of the patient (mean dose 7.85 X 106 per Kg) which decrease to a mean of 5.85 X 106 per Kg post thawing of frozen stem cells. Stem cells viability after thawing ranged from 77 percent to 90 percent of total CD 34+ cells. All the patients had neutrophil and platelet engraftment before they were discharged. Days taken for engraftment of neutrophil ranged from 10 to 15 days (mean 11.5 days) whereas for platelets, it ranged from 14 to 43 days (mean 23.5 days).

Conclusion: Peripheral blood stem cells can be cryopreserved using a solution of 10% DMSO, 20% albumin with 6% HES up till 8 weeks without significant decrease in number and viability of CD34+ cells.

2019 May; 13(Suppl 1): S32–S100.

PP 55: Viability of peripheral blood stem cells after storage at 4°C in liquid state for 72 h

Background: Optimum storage condition with reference to storage medium and temperature has not been clearly defined for non-frozen storage of peripheral blood stem cells (PBSCs). Liquid storage of PBSCs may be indicated in both autologous and allogenic transplantation setting and hence evidence base for the permissible time limit for such storage is need of the hour.

Aims: To study the viability of PBSC during storage in the liquid form at 4 degree celcius for 72 hours post- collection.

Methods: 16 PBSC products harvested for hematopoietic stem cell transplant (5 in Autologous and 11 in Allogenic transplants) were specifically tested for viability by flowcytometry using the 7- AAD protocol within 2 hours of the collection and after 24 hours, 48 hours and 72 hours of the PBSC collection. The viability data was analyzed statistically using paired ‘t’ test.

Results: The mean PBSC viability immediately after the harvest was 97.65%. Viability decreased to a mean value of 96.2% at 24 hours, 93.75% at 48 hours and 88.5% at 72 hours. This reduction in the viability was statistically significant when the baseline viability and 72 hours viability of the PBSC samples were compared. The average reduction in viability over 72 hour's storage was 9.15%. The adequacy of the total PBSC dose was not affected significantly by this reduction in viability. All patients had a satisfactory WBC and platelet engraftment after infusion of the PBSCs in above cases.

Conclusion: PBSC integrity was optimally maintained when they were stored in liquid state at 4 degree Celsius for upto 72 hours after harvest. This can be useful to limit the use of DMSO for cryopreservation and assist in decision making to harvest PBSC's in a donor on Day 3 or Day 4 of G-CSF and further storage till day “0” of patient is attained.

2019 May; 13(Suppl 1): S32–S100.

PP 56: Elucidating neuronal regulation of epithelial stem cells and regeneration in the adult salivary gland: Pilot cell culture study towards development of regenerative therapies for damaged salivary glands

Background: Stem cell therapy can restore salivary function by regenerating acinar cells that are destroyed by radiation for cancers. Parasympathetic innervation and acetylcholine mediated signaling is essential for acinar cell regeneration, via regulation of SOX2+ progenitor cell population in fetal models. Studies in adult models are lacking. This project aimed to study effect of same in regeneration of adult murine salivary glands.

Aim: To demonstrate that parasympathetic nerves and the neurotransmitter acetylcholine/mimetics can support acinar cell proliferation, differentiation and regeneration in adult murine salivary gland.

Methods: A series of experiments involved tissue explant co-culture assay and an organoid assay using neuronal factor carbachol and embryonic parasympathetic ganglia. The explant was prepared from 6 weeks old adult mice and the ganglia were harvested from embryonic mice. The complex interaction between epithelia, parasympathetic nerves and neuronal factors was studied in tissue culture model at different time points (day 2, 3, 6, 9). The immunostaining markers E-Cadherin (epithelial), SOX-2 (progenitor cell), β3 Tubulin (neural tubulin), Caspase 3 (apoptosis), dapi (DNA) were used. Assessment of qualitative/semi-quantitative changes representing acinar cells and progenitor cells was done using confocal microscopy and ImageJ software. The data was analyzed using statistical analysis software (SPSS, IBM corp.). Key parameters were the mean, standard deviation and range for data on cell counts, nuclear size, organoid size and numbers.

Results: Carbachol promotes cell survival and tissue integrity in explant ex vivo culture. Co-culture with embryonic ganglia promotes growth of nerves (β3 Tubulin) and progenitor cell (SOX2) survival. Neuronal factors cause growth, repair and regeneration of acinar epithelia. Carbachol promotes cell survival, adhesion and growth of individual cells in suspension to produce implantable organoids.

Conclusion: Acetylcholine mimetics may be a viable therapeutic to promote SOX2+ mediated acinar cell replacement following injury or disease, thus restoring salivary function to patients.

2019 May; 13(Suppl 1): S32–S100.

PP 57: Evaluating the role of mononuclear cell count in predicting the cell dose of apheresis derived peripheral blood stem cells by correlating with flow cytometry based cd 34 enumeration

Background: The engraftment of transplanted PBSCs is predicted by the enumeration of CD34+ cells, which may have to be carried out multiple times before, during and after the PBSC harvest procedure. Mono-Nuclear Cell (MNC) count of hematopoietic stem cells may be employed for estimation of the yield during mid-cycle of PBSC harvest and provides a low-cost alternative to CD 34 enumeration in such situations.

Aims: To examine the robustness of the correlation between CD34+ cell yields and the MNC counts in PBSC product in autologous and allogenic stem cell transplants.

Methods: The study included 107 consecutive apheresis procedures performed on donors from August 2015 to July 2018. The study group was divided into two categories, comprising of allogenic and autologous harvests. Correlations between the number of CD34+ cells and MNC in the apheresis products were assessed using Karl Pearson's Coefficient of Correlation with Significance levels (p value) set at 0.05, using SPSS-24 Software.

Results: In allogenic donors, the average cell dose of PBSC product in terms of CD 34 + cells was 7.6 X 10e6/Kg recipient body weight and MNC count was 6.14 X 10e8 MNCs/Kg. In autologous donors, the average cell dose of PBSC product in terms of CD 34 + cells was 2.3 X 10e6/Kg and MNC count was 3.9 X 10e8 MNCs/Kg. The correlation between the stem cell product yield calculated using MNC count and CD34 count was found to be moderately significant (0.673) in case of allogenic donors with p value = 0.001, but no statistically significant coefficient of correlation could be observed in case of autologous donors.

Conclusion: The findings of this study suggest that the MNC count can be employed as a useful and cost effective method of predicting the yield during the mid-cycle of the PBSC harvest procedure in allogenic donors.

2019 May; 13(Suppl 1): S32–S100.

PP 58: Analysis on composition of leukapheresis product - A comparison between intermittent and continuous flow apheresis equipments

Background: CD34+ cells possess lymphocyte like morphology and the apheresis equipment ideally harvest in the zone between the platelet layer and granulocyte layer. As the interface between the buffy coat zone and red cell zone is narrow, red cells and platelets are being collected during the leukapheresis procedure. CD34+ cells represent a small proportion of the leukapheresis bag content. The present study aims to analyse the cellular content of leukapheresis product collected by both intermittent and continuous flow apheresis equipment.

Methods: Retrospective analysis involving 111 leukapheresis procedure for 85 patients. Procedures were performed in both intermittent flow (MCS+, n=62) and continuous flow (Spectra Optia, n=49) apheresis equipment. The procedure end point was 2 times the total blood volume processed. The bag sample was aliquoted and tested using LH750 cell counter and FC500 Flow cytometer. The difference between intermittent and continuous flow equipment was compared through ‘t’ test. Linear regression analysis was performed to evaluate the impact of product composition in association between the equipments.

Results: The procedure settings like weight, complete blood count, preCD34 count, %TBV processed was similar between the equipments. There was no significant differences observed in terms of product volume (p=0.67), CD34 count (p=0.7), CD34 dose/kg (p=0.6), MNC dose/kg (0.08) between two equipments. Continuous flow equipment had minimal red cell (Hct 2.5% vs 9.6%, p<0.05), and platelet (1124 vs 1423*10e3/μl; p=0.03) contamination. However, the intermittent flow had better WBC content (255 vs 215*10e3/μl, p=0.04) and thereby increased TNC dose/kg (10.1 vs 7.9 x10e8, p=0.01). The mobilization regimen used did not affect the bag content between the equipments.

Conclusion: The mean red cell and platelet content were 3.8 and 1.3 times higher using intermittent flow apheresis device. Hence patients with relatively low Hb or risk of 2nd procedure, leukapheresis using continuous flow device would be better.

2019 May; 13(Suppl 1): S32–S100.

PP 59: Cost effectiveness of hematopoietic stem cell transplantation compared to transfusion chelation for treatment of thalassemia major

Background: Hematopoietic stem cell transplantation (HSCT) is the only cure for thalassemia major (TM), which inflicts a significant 1-time cost. Hence, it is important to explore the cost effectiveness of HSCT versus lifelong regular transfusion-chelation (TC) therapy.

Aims: This study was undertaken to estimate incremental cost per quality-adjusted life-year (QALY) gained with the intervention group HSCT, and the comparator group TC, in TM patients.

Methods: A combination of decision tree and Markov model was used for analysis. A hospital database, supplemented with a review of published literature, was used to derive input parameters for the model. A lifetime study horizon was used and future costs and consequences were discounted at 3%.

Results: Results are presented using societal perspective. Incremental cost per QALY gained with use of HSCT as compared with TC was Image 17 64,096 (US$986) in case of matched related donor (MRD) and Image 18 1,67,657 (US$2579) in case of a matched unrelated donor transplantation. The probability of MRD transplant to be cost effective at the willingness to pay threshold of Indian per capita gross domestic product is 94%.

Conclusion: HSCT is a long-term value for money intervention that is highly cost effective and its long-term clinical and economic benefits outweigh those of TC.

2019 May; 13(Suppl 1): S32–S100.

PP 60: Technical challenges in ex vivo culture of red cells

Backgroud: Since there is a consistently rising demand for red cells throughout the world, it is imperative to explore safe and reliable alternatives to donor blood. Ex-vivo culture of red cells from haematopoietic stem cells (HSC) has the potential to revolutionize transfusion medicine. However, culture of red cells is not only costly, but fraught with technical challenges.

Aim: To grow ex vivo reticulocytes from HSC (as part of a larger project) and document the challenges in erythroid cell cultures.

Methods: Peripheral blood mononuclear cells were retrieved from 2 platelet apheresis cones of anonymous healthy human platelet donors after informed consent. 95% pure population of CD34+ were obtained using magnetic cell sorting isolation (Milteny Biotech GmbH). Cells were seeded in primary erythroid culture medium (Iscove's Modified Dulbecco's Media, Biochrom) at a concentration of 1 x 105 cells/ml. The cultures were maintained in stationary plastic tissue culture flasks (not followed by mechanical stirring) at a low density of 1-5× 105 cells/mL at 37°C in a humid atmosphere of 5% CO2 in air as per Griffiths et al's (2012) 3-stage protocol. Daily cell counts in duplicate were carried out to study proliferation. Cytospinswere prepared to study cell morphology. On day-21, erythroidcultures (3.5 ml) were filtered to obtain a pure population of reticulocytes.

Results: The cells followed a normal growth pattern until day-7. After that, the cultures entered a premature plateau phase due to low cell density caused by experimental error. Although cells remained viable, proliferation was negatively affected. The doubling time in the exponential stage was around 13 hours. Cytospins prepared from erythroid cultures revealed normal erythroiddifferentiation.

Conclusion: Our cultures had no contamination issues, but sterile and accurate technique are paramount to obtain a good yield of reticulocytes for research and therapeutic purposes. It would be challenging to establish a cost-effective process for large-scale production.

2019 May; 13(Suppl 1): S32–S100.

PP 61: Anaphylaxis to single donor platelet transfusion - A case report with definite imputability and grade IV severity

Background: Platelet transfusions play a decisive role in therapeutic regimens for patients with hematologic/oncologic diseases who develops severe thrombocytopenia. Allergic transfusion reactions are reported to complicate approximately 1-3% of all blood transfusions especially to plasma containing products. These reactions are usually mild and often accompanies cutaneous manifestations. Anaphylactic shock following blood transfusion though rare, but life threatening occurs with a frequency of 1:20000 to 47000. Here we report a case of transfusion reaction to SDP which lead to the death of the patient.

Case Report: A 42 year old female, a known case of Hodgkin's lymphoma, had completed her 6 cycles of chemotherapy was posted for autologous stem cell transplantation (ASCT). She was started on to GCSF on a twice daily basis for 4 days. First collection of stem cell was uneventful with a CD 34 count of 3.13*106 cells/kg. This prompted to perform a second collection for the patient. With regards to the low platelet count 1 SDP transfusion was initiated but afte 3 minutes patient developed severe breathing difficulty, flushing, frothing from the mouth and later went to cardiac arrest. She was resuscitated and intubated. Next day, she was declared death. All transfusion reaction workup in the blood bank was normal. Blood culture reports were sterile, Chest X ray findings and supporting evidences ruled out TRALI and hypotensive reactions. There was no IgA deficiency reported. Patients clinical symptoms, Serum tryptase levels (>200 micrg/dl) suggested the features of mast cell degranulation and hence pointed towards anaphylaxis to platelet transfusion.

Conclusion: Anaphylaxis reactions are mainly caused by the antibodies produced in recipients who have been transfused repeatedly. Though the etiopathology often suggested IgE or anti IgA, release of both preformed granule-associated mediators and newly generated lipid-derived mediators from the platelets contributes to the amplification and prolongation of the reaction.

2019 May; 13(Suppl 1): S32–S100.

PP 62: Assess the serum ferritin in nonremunerated voluntary blood donors and to correlate iron stores with other hematological parameters

Aims: This Study was planned to assess the iron stores in voluntary blood donors and see the effects of frequent blood donation in the development of iron deficiency anaemia.

Methods: A total of eight 84 blood donors participated and informed written consent was taken from all donors. These were categorized into regular blood donors who were further classified on the basis of number of donation in the preceding years and first time donors.

Results: Mean values of first time donors vs. regular donors for Hb (gm/dl), MCHC (%), Serum Ferritin (ng/ml) and Serum Iron (μg/dl) were 15.42 vs. 14.20 (p – 0.23) ; 35.19 vs. 34.45 (p – 0.455) ; 58.94 vs. 32.72 (p – 0.03) ; 87.00 vs. 85.71 (p – 0.19) respectively. It was seen that serum ferritin concentrations were significantly different when comparing first-time donors with regular voluntary blood donors especially groups II & III (58.94 vs. 31.38, p = 0.002 and 58.94 vs. 21.80, p=0.0001). In our study, we also compared ferritin levels in different age groups of first time vs regular blood donors and there were statistically significant variations in 18 - 20 years (57.68 vs. 17.40, p = 0.02) and 31- 40 years (89.36 vs. 35.03, p = 0.001) age groups.

Conclusion: This study shows in comparison to the first time donors, regular blood donors especially in group II & III (donation frequency >2/yr) have more negative iron balance (<30 mg/l) which lead to iron deficient erythropoiesis (MCV<80 fl) followed by iron deficiency anemia. Since with this high frequency of blood donors with iron deficiency suggests a need for a more accurate laboratory trial, since Hb measurement alone is not sufficient for detecting and excluding voluntary blood donors with iron deficiency without anemia.

2019 May; 13(Suppl 1): S32–S100.

PP 63: Role of predonation water intake in donor adverse reactions

Background: Blood donors especially first time donors are more prone for blood donation related adverse reaction and the experience of such symptoms can contribute to a loss in the retention of blood donor. Many studies of water ingestion has shown to increase tolerance for an upright posture, due to sympathetic activation, increase in peripheral resistance and more efficient regulation of cerebral blood flow approximately 30 minutes after ingesting about 500ml of water. Thus, Water ingestion may be sufficient to reduce the risk of syncope related reactions during blood donation by changing hemodynamic responses.

Aim: The aim of the study is to analyze whether pre donation of 500 ml water intake will reduce the syncope related donor adverse reaction among voluntary non remunerated blood donors (VBDs) in the outdoor blood donation camp.

Materials and Methods: Among 600 donors 570 Male and 30 Female voluntary blood donors were randomly assigned into two groups- 300 donors without water intake and 300 donors with 500 ml water intake approximately 30 minutes before donation. Participants have been assessed for syncope related donor adverse reactions during and after blood donation.

Results: In our study total 26 donors reported syncope related adverse reactions. Out of 26 donors with syncopal attack eighteen of them were male and eight were female. Among them twenty two donors (22/300) who haven’t taken water before donation have experienced syncope related adverse reactions and four donors (4/300) who had taken 500 ml water 30 minutes before blood donation reported syncope related adverse reactions.

Conclusion: Result of the present study suggest that predonation water intake of 500 ml water approximately 30 minutes before blood donation may be a simple and cost effective strategy to enhance the blood donation experience and decrease the donor syncope related adverse reactions.

2019 May; 13(Suppl 1): S32–S100.

PP 64: Vitamin B12 level in blood donors

Background: Vitamin B12 deficiency is common in our country as majority of our population is vegetarian or cannot afford regular non-vegetarian diet. We tried to check vitamin B12 level in blood donors who came to donate at our center or outdoor camp.

Aims: To check vitamin B12 deficiency in voluntary blood donors at our center and guide the donors who are deficient.

Methods: We divided the donors into two categories viz. donors rejected due to low hemoglobin and donors accepted (taken as control). We did CBC and carried out peripheral blood smear examination. Depending on the results, we got vitamin B12 assayed (by CLIA) of those donors in whom we suspected vitamin B12 deficiency – either MCV more than 100 fl or peripheral smear examination showed macrocytes and/or hypersegmented neutrophils (≥ 6 lobes) or 5% neutrophils having 5 lobes.

Results: 490 donors were rejected due to low hemoglobin (<12.5 g/dL) in our study. Vitamin B12 was done on 46/490 samples suspected to have deficiency of the same. Only 4 out of 46 had MCV >100 fl (maximum 125 fl). 37/46 (80.43%) or 37/490 (7.55%) had vitamin B12 deficiency. We got vitamin B12 done on 28/425 accepted donors’ samples (hemoglobin ≥12.5 g/dL). 24/28 (85.71%) had normal values while 4/28 (14.29%) had deficiency, overall ratio for deficiency being 0.9%. When this data is applied to whole donor population, 100/490 rejected donors (20%) and 61/425 accepted donors (14.4%) had vitamin B12 deficiency. All the identified vitamin B12 deficient persons were appropriately treated.

Conclusion: Vitamin B12 deficiency is common in the donor population of South Gujarat, however it is significantly higher in rejected donors (X2=5.3 with Yates’ correction, P=0.02). MCV is a poor indicator for B12 deficiency in this voluntary donor population probably because of concomitant iron deficiency from repeated donation.

2019 May; 13(Suppl 1): S32–S100.

PP 65: Single donor platelet donation: Pleasure or pain? - Donor's perspective

Background: Apheresis is a procedure in which whole blood is removed from the body and passed through the equipment that separates out one or more particular blood constituent. In a plateletpheresis procedure, platelets along with a portion of plasma are removed and the remaining constituents are returned to the donor. A plateletpheresis procedure typically takes 45 to 90 minutes.

Aim: To evaluate the subjective perception about single donor platelet donation by apheresis among voluntary blood donors.

Methods: This observational study was carried out in June, 2018 at the Department of Immunohaematology and Blood Transfusion, Sri Ramachandra Medical College and Research Institute. Continuous flow cell separator (COM.TEC) was used for plateletpheresis procedure. Donors who had donated platelets (by apheresis) were asked to fill a post-donation questionnaire and the responses were evaluated (n=20).

Results: All 20 participants were males. Mean age of the participants was 28.6 years (Range: 20-49 years). All participants had donated whole blood earlier. Four out of 20 participants felt anxious/scared after the medical officer described the plateletpheresis procedure/looking at the cell separator. Three out of 20 participants weren’t comfortable during the procedure. All participants considered (a) the separation of particular blood constituent, (b) increased blood donation frequency and (c) decreased donor exposure rate in the recipient as the desirable factors of apheresis donation. The participants considered the effects of citrate toxicity like tingling sensation/twitching (9 out of 20) and the longer time duration (9 out of 20) compared to whole blood donation as the undesirable features of the plateletpheresis procedure. All participants except one had expressed their willingness to undergo this procedure again.

Conclusion: From the donor's perspective, apheresis technology is complex and a donor might be facing some logistical difficulties like increased procedural time. Yet the donor plateletpheresis procedure is accepted well among the blood donors.

2019 May; 13(Suppl 1): S32–S100.

PP 66: Analysis of donor deferral due to low hemoglobin: A 3 years retrospective study

Background: Low hemoglobin is a significant cause of temporary deferral all over the world. All donors are subjected to hemoglobin estimation which should not be less than 12.5 gm/dl. This study identifies the prevalence of donor deferral due to low hemoglobin in a tertiary care hospital in eastern India.

Aims: To analyze and find out prevalence of anemia in prospective blood donors, access the grade of anemia in relation to age and sex and suggest strategy for management of donors deferred due to low hemoglobin.

Methods: This retrospective study was conducted in SUM Hospital Blood Bank, Bhubaneswar Odisha from May 2015 to June 2018. A total of 31,680 donors were screened. Hemoglobin estimation was done by HEMOCUE HB 201+ ANALYSER (Sweden).

Results: A total of 2882 (9.09%) were deferred [Total donor =31,680] due to various reasons. Low hemoglobin accounted for 1245 (43.19%) [males 742 (59.59%) and females 503 (40.41%)] of all deferrals. Donors deferred due to low hemoglobin in age group of less than 20 years were 46 (3.6%), 20-40 years were 939 (75.4%) [males-486 and females 452] and more than 40 years were 260 (20.88%). A total of 585 (78.8%) males had mild anemia in contrast to females 362 (71.96%) while moderate anemia was seen in 157 (21%) males and 141 (28.03%) females. In female donors, 12 (0.96%) had hemoglobin in the range of 12-12.5 gm/dl which doesn’t constitute anemia according to WHO criteria.

Conclusion: Deferral due to low hemoglobin leads to non retention of donors. To avoid non-anemic female donor (according to WHO criteria) deferral, present deferral criteria may be revisited. Prolonging the inter-donation interval and limiting donation to 350 ml to allow recovery of iron stores can be considered in this group of female donors. Focussed counselling and their referral for further investigation and appropriate treatment will help their return to voluntary donor pool.

2019 May; 13(Suppl 1): S32–S100.

PP 67: Physiological and methodological factors affecting hemoglobin estimation in blood donors

Background: Blood donor hemoglobin estimation is an important test that is performed prior to blood donation. It serves the dual purpose of protecting the donors’ health against anemia and ensuring good quality of blood components. Besides the technique, the anatomical source of blood sample, posture of the donor, timing of sample and other biological factors affect the accuracy and reliability of hemoglobin estimation.

Aims: To study the effect of various methodological and physiological factors on the hemoglobin levels of blood donors.

Methods: In this prospective study, 200 voluntary non remunerated blood donors were recruited after obtaining consent. All the demographic details were recorded after obtaining a detailed history and medical examination. 100 samples were obtained in winter months (December and January) and 100 in summer months (May and June). Capillary blood samples were obtained from finger prick and pre-donation venous blood samples were obtained from ante-cubital vein. In 100 donors samples were obtained in sitting position and from other 100 in recumbent position. All the samples were tested with hemoglobinometer (Hemocue 301).

Results: The mean hemoglobin of capillary blood was found to be significantly higher than venous blood. The mean hemoglobin was significantly higher in male donors, donors who were residents of hilly areas and donors with history of chronic smoking. The mean hemoglobin was lower in recumbent position as compared to standing but the difference was not significant.

Conclusion: The source of the blood sample is the most important variable for the accuracy of a technique and critical determinant of donor eligibility in borderline cases. Each blood centre should develop its own policy for borderline hemoglobin results, keeping in view the physiological factors which influence the result of hemoglobin testing.

2019 May; 13(Suppl 1): S32–S100.

PP 68: Comparative evaluation of predonation hemoglobin screening methods

Background: The predonation hemoglobin (Hb) estimation is the only laboratory test done on blood donors to determine an individual's eligibility to donate blood with an intention to prevent bleeding an anaemic donor. With availability of wide range of screening methods, no single technique has emerged as the most appropriate and ideal for screening blood donors. Objective: The primary objective of the study was to compare results of copper sulphate method and haemoglobin meter with the gold standard Cyanmethhemoglobin (CyanHb) method.

Methods: Prospective observational study done in 238 blood donors. Sample analyses were done using Copper sulphate solution, haemoglobin meter and Cyanmethhemoglobin method.

Results: The mean hemoglobin values obtained by Cyanmeth and, Hemoglobin meter were 15.20g/dl, and13.27g/dl respectively. Hemoglobin values obtained by haemoglobin meter and copper sulphate methods were comparable, with sensitivity of 94.25% and 94.36 % respectively.

Conclusion: CuSo4 method and haemoglobin meter can be used alternatively based on resources available. The higher values obtained by CyanHb method could be due to turbidity factor of the lipids in the blood sample.

2019 May; 13(Suppl 1): S32–S100.

PP 69: A study of blood donor deferral due to anemia in the blood bank of a tertiary care centre

Background: Blood donors are deferred from donating blood for several reasons, either permanently or temporarily. Anemia is a major cause for temporary deferral among donors. The aim of our study was to find the incidence of deferred donors due to anemia and to analyse the most common blood group deferred due to it.

Methods: A retrospective study was done from January 2017 to April 2017 in Father Muller Medical College Hospital Blood Bank. Data was collected by reviewing the deferred donor records over four months period. It was analysed by frequency and percentage.

Results: Out of 3354 donors over four months period, 559 were deferred (203 females, 356 males). Of these, 144 were deferred due to anemia (137 females, 7 males). The most common cause of deferral in women was due to anemia, accounting for about 67.48%. It was seen that donors with A Rh positive blood group were mostly deferred.

Conclusion: Donor deferral can be reduced by creating awareness, health education and also by administration of Iron-Folic acid supplementation.

2019 May; 13(Suppl 1): S32–S100.

PP 70: Platelet apheresis and donor safety

Background: Apheresis is generally considered as safe procedure, although limited studies have addressed the issue of donors safety. With regards to the effect of the procedure on hematological indices in these healthy donors.

Aim: Purpose was to study the influence and to identify potential hematologic changes occurring post procedure in first time platelet apheresis donors.

Methods: In this pilot study 35 apparently healthy voluntary adult donors were evaluated. Apheresis procedure was performed on the donors recruited for the first time for platelet donation, using automated cell separator machine. Hematologic measurements (hemoglobin, hematocrit, white blood cells counts and platelet count) were analyzed before and after platelet apheresis in these donors. In addition we also compared the pre and post donation values of Total Serum proteins levels.

Results: We observed a significant drop in hematological parameters following platelet apheresis but without any clinical manifestations of anemia, leukopenia, thrombocytopenia or hypoproteinemia. Decrease found in haemoglobin (p <0.0001), haematocrit (p <0.0001), leukocytes (p <0.0001), and platelets (p <0.0001). On the other hand, we found no significant changes in total serum proteins and also no change in differential granulocytes.

Conclusion: Despite of significant drop in the haematological parameters in both red and white cells, none of our donors manifested symptoms of either thrombocytopenia, leukopenia, anemia or even hypoproteinemia. This study does demonstrate hematological changes post-donation but recommends longterm followup of these donors to ensure complete well being. Larger Multicenter studies are recommended, to help establish actors for donors’ safety in apheresis; This will also bring a major leap in remotivating and repeat recruitment of platelet donors thereby maintaining the safety of the products too; especially given the present trend of double product apheresis collections.

2019 May; 13(Suppl 1): S32–S100.

PP 71: Donor deferral pattern in a tertiary care hospital, Jamnagar, Gujarat, India

Background: Donor selection is must and one of the most crucial step, useful in providing TTI safe blood to the needy patients for transfusion purpose without ignoring the donor's safety.

Aim: (1) To recruit a healthy donor pool for the purpose of safe blood transfusion. (2) To study the incidence and pattern of donor deferral. (3) To implement better recruitment & retention strategy which help in reducing the incidence of the same in the institute.

Methods: A retrospective study over 18 months has been done from January ‘2017 to June’2018 on causes of donor deferral in the IHBT department of Shri M P Shah Govt. Medical College, Jamnagar, Gujarat, India.

Results: A total of 35545 donors were reported between January ‘2017 to June’2018. Total 838 (2.36%) donors were deferred for different reasons which included temporarily deferred 818 (97.61%) & permanently deferred 20 (2.38%) as per the guidelines given by NABH. Out of 35322 registered male donors 743 (2.10%) & out of 223 registered female donors 95 (42.6%) donors were deferred respectively. Major cause of temporary deferral in both the sexes was anemia (62.24%). Other causes included like antibiotic therapy (4.27%), tattoo (5.50%), menstruation (1.83%), alcohol abuse (1.83%) etc. In permanent deferral causes were heart attack (15%), hypo/hyper thyroid (15%), history of convulsion (10%) etc.

Conclusion: (1) From 509 Anemic, otherwise healthy deferral donors 28.09% had low Hb between 12.0 to 12.04 gm/dl which indicates to pay attention on the deferral criteria for the same. (2) Creating awareness on nutritional therapy, alcohol abuse, unreasonable antibiotic use & special awareness in females regarding blood donation.

2019 May; 13(Suppl 1): S32–S100.

PP 72: Evaluation of various methods for hemoglobin screening of blood donors

Background: Despite the wide range of methods available for measurement of hemoglobin, no single technique has emerged as the most appropriate and ideal for a blood donation setup.

Aim: To compare the various methods of hemoglobin screening for their utility and cost effectiveness.

Methods: A prospective study utilizing 200 blood samples was carried out in a blood donation setting for quality evaluation of four methods of hemoglobin estimation: Hematology cell analyzer, Beckman Coulter LH 750 (reference), HCS, HemoCue 201+ and Compolab TS.

Results: Mean value of HemoCue (mean ± SD = 14.05 ± 2.20 g/dl) was higher by 0.4 compared to reference (mean ± SD = 13.65 ± 2.14 g/dl) but not statistically significant (P > 0.05). HemoCue proved to be the best technique (sensitivity 99.2%) whereas HCS was most subjective with 25.2% incorrect estimations. Compolab was almost as sensitive as Hemoque w.r.t Hematology Analyzer.

Conclusion: HCS method gives accurate results, if strict quality control is applied. HemoCue and compolab are more accurate but expensive.

2019 May; 13(Suppl 1): S32–S100.

PP 73: Adverse donor reactions to whole blood donation: A retrospective study of 56,726 blood donors at sms medical college Jaipur

Background: Blood is the most precious gift that one human being can give to another. Blood donation is a safe procedure. Most donors tolerate giving blood very well but occasionally adverse reaction may occur during and at the completion of blood donation.

Aim: To study and analyze the spectrum and prevalence of adverse reactions in whole blood donors.

Methods: This is a retrospective observational type study conducted from 1 June 2017 to 31 May 2018 at Dept. of IHTM, SMS Medical College Jaipur.

Results: The prevalence of adverse donor reactions was (635/56726) 1.12 %. This occurred more frequently in female donors as compared to male donors. The spectrum of adverse donor reactions included mild vasovagal (82.68%), moderate vasovagal (13.07%), Hematoma (2.519 %) and Numbness/Tingling/soreness of arm is 1.732%.

Conclusion: Vasovagal reactions were the most frequent adverse phenomenon in this study. This study reinforces that blood donation is a very safe procedure which could be made even more adverse reaction free with friendly and tactful practices.

2019 May; 13(Suppl 1): S32–S100.

PP 74: Association of reactive thrombocytosis with microcytic hypochromic anemia - A case control study

Background: Safety of blood donors is the prime priority in blood banking and risk of iron depletion in repeat donors is a challenge encountered. Iron deficiency (ferritin <12 ng/ml) is detected in a few repeat blood donors especially females. Currently donor haemoglobin estimation is the only method used by blood bank to protect the donors from ill effects of iron depletion. An association between thrombocytosis and ID has been studied by various authors considering it as a surrogate marker for IDA. Elevated platelet count in ID may also carry the risk of thrombosis as higher aggregable platelets are produced. There are case reports linking ID-related thrombocytosis and thrombosis, and higher incidence of ID in patients with cerebrovascular insult. We attempted to compare the platelet count (PLC) in patients with hypochromic microcytic anaemia (HMA) and without HMA.

Aim: To compare the platelet count between individuals with HMA and without HMA.

Methods: Case control study from 01/04/2018 to 30/06/2018. Cases (n=200) were those diagnosed in clinical pathology laboratory as HMA using peripheral smear, CBC and controls (n=200) with no evidence of HMA in smear and normal haemoglobin. PLC was compared using independent t test.

Results: Mean age of cases were 44.61 years, SD22.07 and that of controls were 31.42 years, SD23.72, 28% of cases and 46.5% of controls were males, Hb values of cases ranges from 2-12.3 g/dL (mean 7.94 g/dL) and that of controls ranges from 10.6-21.6 g/dL (mean-13.9 g/dL). PLC of cases ranged from 15000 to 2,48,000 (mean3.08 x 106/μL)and that of controls ranged from 2600 to 150000 (mean -2.68 x 106/μL). PLC was significantly higher in subjects with HMA (t-2.36, p-0.01). On chi square test, significant association was found with elevated PLC (PLC>4.5x106/μL) and HMA, Odds ratio- 5.448, CI 2.46 -12.03, p-0.000.

Conclusion: An association between IDA and reactive thrombocytosis is detected in the study. While screening donors for whole blood and plateletpheresis procedures, coexistence of these two conditions should be borne in mind.

2019 May; 13(Suppl 1): S32–S100.

PP 75: Plateletpheresis donor deferral characteristics at a tertiary care hospital of eastern India

Background: SDP is utilized as a major transfusion support in critically ill thrombocytopenic patients having bone marrow transplantation, hemato-oncological disorders, critical care patients and in some emergency situations like dengue hemorrhagic fever, active bleeding etc.

Aim: The aim of this study was: (1) To have a background profile of SDP donors and identify some common reasons of deferral. (2) To encourage and retain a pool of voluntary SDP donors.

Methods: This prospective observational study was carried out in the department of IHBT, Medical College Kolkata over a period of one year (jan’17- dec’17). A detailed history was taken and the donors were selected as per the departmental SOP for plateletpheresis donor's selection: weight >55 kg, age 18-60 years, atleast 3 months from last WB donation or 48 hrs from last SDP donation, ABO identical donor., adequate venous access, hemoglobin >12.5 g/dl, platelet count > 150*103/μl., no consumption of NSAIDs for the last three days, non-reactive for TTI markers.

Results: A total of 849 donors were screened for 145 procedures. 91.72% (133) of these were planned procedures and the rest 8.28% (12) were emergencies. Among 849 donors, 449 donors (i.e. 52.88%) were deferred due to various reasons. The commonest cause of donor deferral included poor venous access (n=186; 41.42%), followed by low platelet count (n=119; 26.5%), low body weight (n=59; 13.14%), non-identical blood group with recipient (n=48; 10.69%), low hemoglobin (n=6; 1.34%), others like H/O drug intake, illness etc. (n=28; 6.23%), the least common being TTI reactivity (n=3; 0.67%). A total of 42% (189) were deferred permanently.

Conclusion: SOP modification related to ABO compatibility in donor selection may reduce donor deferral rate. Donor deferred due to temporary causes must be adequately counseled to encourage them for future donations.

2019 May; 13(Suppl 1): S32–S100.

PP 76: Attitude of voluntary blood donor towards repeat and regular blood donation – A prospective study

Background: Self sufficiency and safety in blood is essential to strengthen the health system. The NACO goal is to obtain all blood supplies from voluntary unpaid donor. Continuous efforts are needed for successful motivation and retention strategies.

Aim: To ascertain the attitude of Voluntary Blood Donors towards repeat and regular blood donation.

Methods: During the study period between May and July 2018 the first-time voluntary blood donors who had donated blood in the blood donation camps were given a structured questionnaire to find out the attitude towards repeat and regular voluntary blood donation. These camps were organized following predonation motivation and education about voluntary blood donation by utilizing Information, Education and Communication materials such as projection of documentary film, posters, pamphlets and lecture followed by interactive sessions.

Results: In this study, out of 1003 donors screened, 983 (98.11%) were males and 19 (1.89%) were females. Of the total, 713 (71.3%) donations came from voluntary blood donors and the remaining 290 (28.7%) from replacement blood donors. Overall, 47.8% (478) of donors came from the age group 18-29 years, 32.8% (328) from 30-39 years age group, 16.2% (162) from 40-49 years age group, and 3.4% (35) from those between 50-59 years of age group. Among them, 76% (762) are first time donors and remaining 24% (241) are repeat donors. After knowing the importance of blood donation and the experience gained during the time of first time blood donation, 85% (648) of the first time donors showed their willingness for repeat and regular donation.

Conclusion: Since our voluntary blood donation camps are well-organized according to the scheduled protocol starting from predonation motivation and, education about voluntary blood donation, 85% of the first time donors expressed their willingness for repeat and regular blood donation. Donor education, motivation and experience during first time donation play a vital role in donor retention.

2019 May; 13(Suppl 1): S32–S100.

PP 77: Notification, donor response and counselling of seroreactive voluntary blood donors – A retrospective study

Background: Unsafe blood remains a major threat for the spread of transfusion-transmitted infection (TTI). As per the NACO guidelines, all initial seroreactive donors who had given consent shall be recalled, offered post donation counselling and referred to appropriate facility. Results shall not be informed over phone. Initial response of the donors is unpredictable. However, it is the responsibility of blood transfusion service providers to inform such donors for appropriate counselling and direct them for further management.

Aim: To evaluate notification, response and counselling rate of seroreactive donors.

Methods: This study is an retrospective study performed in The Tamil Nadu Dr. M.G.R. Medical University, Department of Transfusion Medicine over a period of 3 years from 2015 to 2017.

Results: During the study period, out of 5741 blood units collected, 62 donors were seroreactive for HIV, HBsAg, HCV and VDRL respectively were two (2/62), forty three (43/62), fifteen (15/62) and two (2/62). The trained blood bank counsellors could able to notify 52 (83.9%) seroreactive donors over telephone, and the response rate was 92.3% (48/52). For the remaining 10 (16.1%) donors, notifications were sent by post. None of them responded for counselling. Among the 48 responded seroreactive donors for HIV, HBsAg, HCV, and VDRL respectively were two (2/48), thirty three (33/48), eleven (11/48) and two (2/48). They were counselled and referred to integrated counselling and testing centre, Medical Gastroenterology and Venereology departments respectively. The fourteen non-responders were HBsAg (10/14) or HCV (4/14) seroreactive.

Conclusion: In our study, around 16% of seroreactive donors could not be traced. The plausible reason could be due to a migratory population in metropolitan cities. If complete demographic details such as unique identification number (Aadhar Card), Voter's Identification Number, Driving Licence Number, etc., are included, it would increase the chances of donor traceability further.

2019 May; 13(Suppl 1): S32–S100.

PP 78: Effect of predonation intake of water and oral rehydration solution on donor adverse reactions – A comparative study

Background: The first time donors are susceptible for donation related adverse reactions (dizziness, syncope) and it can affect donor retention. Physiologic strategies may reduce an individual blood donor's risk of a reaction, such as predonation intake of water.

Aim: The aim of our study is to compare the effects of pre-donation intake of water and ORS in reducing syncope related adverse reactions among first timevoluntary blood donors.

Methods: The study was undertaken in the Department of Transfusion Medicine, The TN Dr. MGR Medical University during the period from June to August 2018.300 donors (240 Male and 60 female) were randomly assigned into 3 groups: Group A - 100 donors without fluid intake, Group B - 100 donors with 200ml water intake and Group C – 100 donors with 200ml ORS intake approximately 30 minutes before donation. Donors were assessed for adverse reactions (dizziness, syncope). Post-donation instructions were given. Results were recorded in donor questionnaire form.

Results: Our analysis revealed that incidence of syncope related adverse reactions in Group A is more than in group B and group C. Among group A six (6%) donors experienced syncope related adverse reaction when compared to two (2%) donors in group B and two (2%) donors in group C. Out of 10 donors with syncopal attack, 8 were male and 2 were female. Group B and Group C donors who experienced adverse reactions had taken food more than 4 hours before donation. Donors with syncope were managed with minimal resuscitation.

Conclusion: We conclude that predonation oral fluid intake (water or ORS) reduces vasovagal reactions. Results of present study suggests ORS improves donor experience and there is no significant difference in role of water and ORS in reducing VVR.

2019 May; 13(Suppl 1): S32–S100.

PP 79: Awareness of blood donation among 1st year undergraduate college students in Chennai

Background: Blood is a major vital component of human beings. Unfortunately we dont have any chemical equivalent to blood. So blood donation has become essential for the survival of the society.

Aim: Objective of the study is to analyse the knowledge of voluntary blood donation among first year college students.

Methods: A questionnaire prepared by The Tamil Nadu Dr. M.G.R. Medical University was given to the 500 first year college students during voluntary blood donation camps, from the period of June to August 2018. A briefing was given to participants about the objective of the study and assured confidentiality.

Results: In our study among 500 1st year college students who had voluntarily come to donate blood in camps, 86% were aware of eligibility criteria of blood donation, 72% were aware of screening of HIV before transmission, 61% aware of various contraindications for blood donation, 45% aware about duration of blood donation, 40% had misconceptions about blood donation.

Conclusion: Our study reiterates the importance of educating the potential donors who enters in to the eligible age of voluntary blood donation, to maintain and expand the donor pool; thereby countrys blood need is consistently maintained.

2019 May; 13(Suppl 1): S32–S100.

PP 80: Donor hemovigilance: A retrospective study

Background: A safe and adequate blood supply depends on healthy, altruistic volunteers who are willing to donate blood without expecting personal gain despite the risk of discomfort or adverse reaction. Donor “hemovigilance” includes all activities that contribute to improve the healthy outcome for blood donors as well as safety and effectiveness of blood donation process. Usually, blood donation is a safe procedure. However, a small percentage of donors may experience an Adverse Event.

Aim: A Retrospective study was carried out among the voluntary blood donors (VBDs) to estimate the incidence, risk factor, the type of adverse events in whole blood donation process and to provide evidence based support for improving the same.

Methods: This study was done from April 2014 to march 2017 among the VBDs in camps conducted by DTM, TNMGRMU, who fulfilled the eligibility criteria as per the guidelines of DGHS. All the adverse events were investigated.

Results: During the period 117 camps were conducted and 6566 donors were bled. Out of these, 67 (1%) donors had Vaso-Vagal Reaction (VVR) and 25 (0.38%) donors had hematoma. Out of 67 donors with VVR, 19 (28.3%) were females and 48 (71.7%) were males. Incidence of adverse events in our study was 1.38%. Donation-related VVR is a multi-factorial response primarily determined in first-time donors. Among the females Anxiety 9/19 (47%), low body weight 7/19 (37%), painful phlebotomy 2/19 (11%) and increased environmental temperature 1/19 (5%). In males Anxiety 32/48 (67%), low body weight 8/48 (17%), painful phlebotomy3/48 (6%) and increased environmental temperature 5/48 (10%). The cause for hematoma in 5/25 (0.08%) donors was due to inexperienced phlebotomist and others 20/25 (0.30%) the most probable reason would be due to noncompliance of appropriate instructions of arm movements during the procedure.

Conclusion: The knowledge of donor adverse events and the probable risk factors would make blood donation process a pleasant experience leading on to better donor retention.

2019 May; 13(Suppl 1): S32–S100.

PP 81: Assessment of sociodemographic profile and risk factors in human immunodeficiency virus sero reactive blood donors in a tertiary care centre of northern India

Background: The rate of transfusion transmitted Human immunodeficiency virus infection (HIV) has shown a declining trend, but it still remains a major public health issue. This study examined socio-demographic characteristics and risk factors of HIV seroreactive blood donor.

Methods: This retrospective study was conducted in a tertiary care hospital over a period of 18 months. Blood donor samples were screened for HIV using fourth generation enzyme linked immunosorbent assay. The seroreactive donors were notified telephonically and through letters. Responders were counseled and referred to ICTC. Reactive donors not responding to three telephonic calls and letter were labeled as defaulters. Data regarding socio-demographic characteristics, and other relevant history was obtained from donor record.

Results: Of total 32852 blood donations, twenty (0.061%) donations were seroreactive for HIV, nineteen were male and one was female. Eleven of these twenty donors donated first time and nineteen were voluntary. Written correspondence could be sent to nineteen donors (95%) whereas one donor (5%) could not be informed due to incomplete address on donor record card. Telephonic calls could be made to seventeen donors (85%) while three donors (15%) did not respond to calls. Of the fourteen donors referred to Integrated Counseling and Testing Centre (ICTC), nine male donors (47%) revealed history of high risk behavior and one donor (5.2%) was aware of his reactive status on post donation counseling. Three donors (15.7%) gave history of prolonged hospitalization and history of blood transfusion was present in one case. Four donors (21%) had history of tattooing and ear piercing.

Conclusion: The present study stresses the need to strengthen pre donation screening, counseling and to repeatedly track these seroreactive donors so as to eliminate them from the safe donor pool.

2019 May; 13(Suppl 1): S32–S100.

PP 82: Study of adverse events in whole blood donors and its impact on blood donor return rate

Background: Adverse Event (AE) is defined as the symptoms/signs of donor discomfort of sufficient severity, that either the donor called for attention or noticed by the staff. These AE can negatively impact the Blood Donor Return Rates (BDRR) and decrease donor retention.

Aim: (1) Assess the frequency and type of AE, and analyse its relationship with the BDRR. (2) Identify measures to increase the Donor Return Rate.

Methods: A Retrospective single centre study, conducted from January 2015 to August 2018, in the Transfusion Medicine department of a tertiary care hospital. All Whole Blood donations received at the centre and at its outdoor camps were analysed. All AE (systemic and local) occurring during/end of the donations were noted. All donors were followed up and interviewed at 1 month after donation for any delayed complications. Collected data was analysed to determine the frequency of various reactions. Also, a cohort of first time donors in 2015 was created and followed up to see if they return for further donations and the BDRR were calculated.

Results: Total of 93,594 voluntary whole blood donations were received during the study period. 5.19% (4866/93,594) of total donations were complicated by Adverse Events during the study period. Vasovagal reactions accounted for 74.5% of all AE affecting 2.9% of the donors. (2714/93,594). Donations made by first time donors were 56% in 2015 of which 78% donors returned during the study period following an uncomplicated first donation, but only 35% returned following an AE. Of the total donors that did not return during the study period, 91.2% had suffered from some AE.

Conclusion: Vasovagal reactions were the most common AE recorded. Reduced BDRR seen following AE during donation. A robust hemovigilance programme and measures to decrease AE eventually helps increase donor retention.

2019 May; 13(Suppl 1): S32–S100.

PP 83: Frequency of Rh D variants in healthy blood donors and patient population at a tertiary care hospital in south India

Background: The Rh blood group discrepancies may arise when an individual is a variant of D antigen. They are reported as RhD negative or positive depending on the type of reagents and techniques used. RhD variants (weak D, partial D and DEL) should be identified as they may produce anti-D when transfused with normal D positive red cells. Thus, knowing the correct RhD status is essential in antenatal patients, donors, and recipients of blood transfusion.

Aims: The main objective of the study is to find the prevalence of RhD variants and to characterize them by molecular methods in a tertiary care hospital in south India.

Methods: Blood grouping was performed on 9279 blood samples collected from donors and patients of tertiary care hospital in South India. All the Rh negatives and the RhD discrepant samples were then tested with various anti-D reagents by different techniques. Adsorption elution technique was performed to identify DEL variants. RhD variants thus identified were characterized by multiplex PCR, QMPSF, and sequencing.

Results: Among blood samples tested, 6.3% individuals had RhD negative phenotype whereas thirteen samples showed discrepant RhD typing result. Rh phenotype of these samples was CcDee (10), ccDEe (2) and ccDee (1). DEL variant was not identified. Molecular characterization of RhD variants showed Exon 3 duplication responsible for the formation of RhD variant phenotype in approx. 60% cases. Mutations causing Weak D type 25, type 15 and RHD (T201R)-CE(5)-D(1342T) were also identified. No exonic mutation could be detected in two RhD variant samples.

Conclusion: Prevalence of RhD variants in South Indian population is 0.14%. Exon 3 duplication is the most common cause of RhD variant phenotype. Due to high immunogenic nature of RhD antigen, it is important to give correct RhD status to an individual in order to avoid future risk of alloimmunization.

2019 May; 13(Suppl 1): S32–S100.

PP 84: Suspected anti-g antibody in a pregnant women: Report of two cases

Background: Anti-G is one of the cause of Hemolytic Disease of the Fetus and Newborn (HDFN), either alone or in association with anti-C, anti-D or both. The antibody mimics a pattern of anti-C and anti-D reactivity in identification panel and is often present with one or both these antibodies. Differentiation amongst all three antibodies in routine pretransfusion workup is essential in antenatal cases.

Case Report: We report 2 cases where either anti-G or combination of anti-C & anti-D is suspected on advanced immunohematology workup. The HDFN can be largely attributed to the probable presence of anti-G in both the cases. Confirmation of the antibody can be done by adsorption and elution methods.

2019 May; 13(Suppl 1): S32–S100.

PP 85: Distribution of Rh and kell (K) blood group antigens among blood donors in a tertiary care hospital

Background: Red cell antigen phenotyping is very important for minimizing the risk of alloimmunization and haemolytic transfusion reactions.

Aim: To study the pattern of Rh (D,C,E,c,e) and Kell (K) blood group antigen distribution among blood donors of different ethnic groups in our tertiary care hospital.

Methods: This was a prospective observational study carried out over a period of 01 year in SMGS Hospital Jammu. It comprised of voluntary and replacement donors belonging to different ethnic groups i.e Dogras, Gujar Muslims, Non-Gujar Muslims, Kashmiri Pandits (KPs), Sikhs and Christians.

Results: 500 blood samples were collected. Out of these, 420 samples were antigen typed by conventional tube technique and 80 samples were antigen typed by column agglutination technology. As per ethnicity, maximum donors were Dogras (74%) followed by Non-Gujar Muslims (9.4%), Gujar Muslims (9%), Sikhs (5.6%), KPs (1.4%) and Christians (0.6%). RhD positive was found among (94.4%) donors and RhD negative among (5.6%) donors. Among other antigens of Rh system ‘e’ antigen was highest (97.2%) followed by ‘C’ (85.2%), ‘c’ (64.0%) and ‘E’ (20.6%). Frequency of ‘D’ antigen was highest among Sikhs, Christians and KPs (100%) followed by Dogras (95.1%). ‘C’ antigen was highest among Dogras (87.29%) followed by Sikhs (85.71%). ‘E’ antigen was highest among Christians (33.33%). ‘c’ antigen was also highest among Christians (100%) and ‘e’ antigen was highest among Dogras (98.1%). 08 Rh phenotypes were found R1R1 (DCe/DCe) has the highest frequency of (36.0%) and least common phenotype found was r’r (dCe/dce) 0.6% in our study. Frequency of ‘K’ antigen positive donors were 2.6% and was highest among Gujar Muslims and Non-Gujar Muslims (4.25%) followed by Sikhs (3.57%) and Dogras (2.16%).

Conclusion: Distribution of blood group antigens among different ethnic groups can help in creation of donor data bank, for preparing in house cell panels and providing compatible blood for patients with multiple alloantibodies.

2019 May; 13(Suppl 1): S32–S100.

PP 86: Anti-E alloantibody: Its significance in a case of anemia

Background: Rh antigens highly immunogenic. Majority of Rh antibodies of IgG type. Alloantibody induced hemolytic anaemia due to anti E IgG antibody binding to red blood cell surface antigens characterized by extreme hemolysis, typically extravascular. Anti-E hemolytic disease also be strongly considered in differential diagnosis as among most common causes of severe neonatal hemolytic disease.

Aim: To diagnose and treat cause of sudden severe anemia in female aged 35 years.

Methods: 35-year-old female patient started developing fever, generalized weakness, loss of appetite, nausea, vomiting, jaundice, anemia progressing to pancytopenia, splenomegaly. Continuous fall in hemoglobin. Also continuous evidence of extravascular hemolysis in spleen, since last 15 days. Hematuria noted after blood transfusion. Peripheral smear showed anisopoikilocytosis, spherocytosis and pancytopenic picture. Reticulocyte count on lower side (0.5%). Forward reverse blood grouping showed no group discrepancy. Direct antiglobulin test DCT performed on patients blood sample revealed positive (1+) agglutination with polyspecific antihuman globulin (anti-IgG and anti-C3d). ICT, Indirect Coombs Test, positive 2 +(Antisera make Tulip Diagnostics). Patient's serum tested against cell panel with screening red cells using gel technology (LISS-Coombs Card, Ortho Biovue System, Ortho Clinical Diagnostics), reacted against cells containing E antigen and behaved as an incomplete antibody. Identification panel (Ortho Biovue System, Ortho Clinical Diagnostics) confirmed presence of anti-E antibody with titer of 1:8 (tube method). Auto-control negative. Eluate tested using commercial acid elution kit (Ortho Clinical Diagnostics), confirmed specificity to be anti-E. Extended red cell typing of patient cells negative for E antigen. Di-thiothreitol treatment of serum confirmed presence of IgG type of antibody. Enzyme treatment showed increased reactivity.

Results: Patient diagnosed as a case with alloantibody anti E.

Conclusion: Detecting blood antibodies: auto, allo, cold or warm, important. Role of Transfusion Medicine laboratory in diagnosis of patient with hemolysis and anemia emphasised.

2019 May; 13(Suppl 1): S32–S100.

PP 87: Cutting edge technology in immunohaematology – Responsive automation

Background: In the field of Transfusion Medicine as elsewhere, there is a constant demand for maximum efficiency with optimal turnaround times and minimum staff. Fully automated immunohematology analysers are now being used by many transfusion services to optimize laboratory efficiency. Some of these analysers have what is known as “Responsive Automation” that brings with it a “transformational” approach to immunohaematology testing.

Aim: Evaluation of the various applications of responsive automation in avoiding errors, standardising techniques and improving turnaround times in the blood bank.

Methods: Samples, run in the analyser over a period of five months were analysed using real time alert remote connectivity. Tests included patient and donor groupings, antibody screens, cross matches and titrations. Some of the parameters analysed were the average turnaround time, first pass yields, reagent usage and top error codes.

Results: The use of responsive automation gave us an insight into the day to day operations of the blood bank, including the turnaround times and pass yields. This function of the analyser has helped us resolve diagnostic checks throughout sample processing since the remote connectivity technology monitors every critical step in the automation process. The level of automation provided permitted remote logins and the ability to review results using the advanced image verification system. Time spent by the technical staff on other jobs like planned maintenance could be audited with the above facility and reasons for delays in this process could be analysed and corrected. This, in turn, also contributed to the optimisation of the turnaround times in our blood bank.

Conclusions: Responsive automation and remote connectivity provide solutions for better laboratory efficiency.

2019 May; 13(Suppl 1): S32–S100.

PP 88: “A end” subgroup: A case report of a rare subgroup of A

Background: A end is a weak subgroup of Blood group “A”, found rarely in general population. It is not detected by routine forward and reverse blood grouping but detected by Adsorption/Elution technique along with saliva testing for A, B and H antigens. Although it is subgroup of “A” but it lacks “A” antigen in saliva and contains only “H” antigen.

Case History: A 46 years male patient with fracture right femur was admitted in the Department of Orthopedics, AIIMS Raipur (C.G.). He had no significant past history including any previous blood transfusion or any alloantibody identification. He had no significant family history. The Department of Transfusion Medicine & Blood Bank received the blood sample of the patient for cross matching. On blood grouping, the patient's sample showed 1w+ agglutination & turbidity on forward grouping with anti-A, which came to be mixed field reaction on microscopy.

Methods: The sample was further tested by Adsorption and Elution procedure for confirming weak subgroup of A or B, which showed it to be a weak subgroup of A. It showed a strong positive reaction with anti-H. Further saliva testing was done to which determined presence of only H substance.

Results: Only H substance was detected in saliva sample. Adsorption/Elution confirmed it to be a weak subgroup of A, and Saliva testing confirmed it to be the weak subgroup of A having only H substance in saliva. Also, a mixed field reaction was seen with anti-A & anti-AB antisera.

Conclusion: As the best possible subgroup showing this reaction pattern could be A end, so it was concluded to be A end subgroup. Identification of the rare subgroups is important because this may be mistyped as group O, if transfused with O group unit can show decreased red cell survival or delayed hemolytic transfusion reaction.

2019 May; 13(Suppl 1): S32–S100.

PP 89: Prevalence of A1 and A2 along with anti-A1 in donor and patients in a tertiary care centre

Background: Among A and AB blood groups, A1 & A2 are major subgroups. A1 subgroups are major subgroups which constitutes around 80% and 20% are A2 approximately. Sometimes anti-A1 present in A2 serum becomes clinically significant when they react at 370 C and causes hemolysis of donor red cells.

Methods: Blood grouping was done with standard Test Tube technique using antisera of Tulip Diagnostics and in-house pooled suspension. A1 and A2 were distinguished on basis of agglutination reaction with anti-A1 lectin (Tulip Diagnostics). A2 serum was tested with A1 red cells at room temperature, 370 C for presence and clinically significance of anti-A1.

Results: A total 1058 blood samples of both patients and donor were analyzed with A and AB blood group 76.37% and 23.63% respectively. Among A blood group, 96.78% were A1 where as 3.22% were A2. While among AB blood group prevalence of A1B and A2B were 90.8% & 9.2% respectively. Out of 547 patients A1 and A2 were 97.3% and 2.7%, while A1B, A2B sub types were 90.65% and 9.35% respectively. Of 533 donors prevalence of A1, A2 and A1B, A2B were 96.38%, 3.62%, 91.6% & 8.4% respectively. Out of 13 A2B patients 3 (23.07%) showed presence of anti-A1 antibody.

Conclusion: Although the presence of clinicaly significant anti-A1 antibody is rare which causes hemolysis of donor A1 red cell but we suggest testing for anti-A1 in all patients of A1 subgroup before transfusion of blood.

2019 May; 13(Suppl 1): S32–S100.

PP 90: Prevalence of direct antiglobulin test positivity in oncology patients at the tertiary cancer care center

Background: The DIRECT ANTIGLOBULIN test (DAT) is a simple test used to determine if red cells have been coated in vivo with immunoglobulin, complement, or both. A high prevalence of DAT positivity in patients with mainly hematological malignancies has been reported.

Aim: To study the prevalence of DAT positivity in oncology patients.

Methods: In this retrospective study the DAT test requests from June 2016 to June 2018 were studied. The DAT testing was done using EDTA samples, done immediately on receipt of the samples by Column Agglutination Technology.

Results: Total of 154 oncology patients were tested for DAT. Out of this 30 (19.5%) were positive for DAT and 124 (80.5%) were Negative for DAT. Total male patients were 82 (53.2%) of whom 11 (13.4%) were positive and female patients were 72 (46.8%) of whom 19 (26.4%) were positive. Blood group wise prevalence showed 49 (31.8%) of O Positive, 7 (4.5%) of O Negative, 48 (31.2%) of B Positive, 4 (2.6%) of B Negative, 27 (17.5%) of A Positive, 4 (2.6%) of A Negative, 14 (9.1%) of AB Positive and 1 (0.6%) of AB Negative. Distribution of patients based on age group between 1-10 were 4, between 11-20 were 12 (25% positive), between 21-30 were 13, between 31-40 were 23 (26% positive), between 41-50 were 32 (18.6% positive), between 51-60 were 27 (18.5% positive), between 61-70 were 32 (21.9% positive), between 71-80 were 11 (27.3% positive). The patient with hematological malignancy were 116 (17.2% were positive) and solid organ tumors were 38 (26.3% were positive).

Conclusion: Clinical considerations together with laboratory data should dictate the extent to which a positive DAT result is evaluated. These autoantibodies can be of IgG, IgM, or IgA isotypes. The pathologic and clinical features of AIHA relate to the autoantibody class, thermal amplitude, and their efficiency of inactivating complement.

2019 May; 13(Suppl 1): S32–S100.

PP 91: Optimization of blood safety through essential characterization of naturally occurring lewis antibody

Background: Lewis antibodies are usually naturally occurring however they may have clinically significant, IgG type and may cause haemolytic transfusion reaction. The present study depicts the clinical significance and detailed characterization of Lewis antibodies in blood donors and patient populations and their implication in safe blood transfusion.

Aims: Aims of our study was to observe the clinically significant Lewis antibodies in Eastern India populations.

Methods: The prospective study included 7 individuals who were detected with Lewis antibodies. Further investigations were performed for detailed characterization of these antibodies with regards to antibody type, thermal amplitude, titre and enzyme study and secretor status of the individuals.

Results: Of the 69354 donors and patients subjected to antibody screening, Anti-Lea was detected in 7 with none having anti-Leb. All showed the Le (a-b-) phenotype with 4 presenting with IgG anti-Lea optimally reacting at 37°C with a highest titre of 32. Where all 7 were ABH secretors however none revealed any Lewis substances. For patients requiring transfusion compatible Lea antigen negative red cell units were issued without any adverse events.

Conclusions: As naturally occurring Lewis antibodies may be clinically significant and cause haemolytic transfusion reaction therefore identification and detailed characterization of antibody in blood donor or recipient is very crucial to blood safety.

2019 May; 13(Suppl 1): S32–S100.

PP 92: Serological evaluation of subgroups of ‘A’ and ‘AB’ in blood donor population in eastern India

Background: Subgroup of ‘A’ was first described by von Dungern in 1911. Broadly such subgroup includes A1, A2 and weak A subgroup. Similarly AB subgroup includes A1B and A2B. Detection of weak A subgroup such as A3, Am, Aend, Ay and Ael are routinely not performed in the blood bank. Such serological investigations are complex, time consuming and require technical skill. Ours being a referral immunohematology laboratory we perform serological evaluation of ‘A’ and ‘AB’ subgroups.

Aims: The aim of this study was to observe the frequency of A and AB subgroups in the blood donor population of Eastern India.

Methods: Over a period of five years donor blood group was typed using column agglutination technique and standard tube technique. Anti A1 lectin study was done for all donors with group A and AB. Subgroup of A and AB was confirmed by serological reactions defined by previous authors. Weak subgroups were determined by adsorption elution technique and saliva inhibition test.

Results: A total of 36789 donors were typed for ABO and Rh group. Among 8941 group ‘A’ donors A1, A2 and weak subgroups were 7358 (82.3%), 1534 (17.2%) and 4 (0.05%) respectively. The weak subgroups serologically investigated were Ay (n=2), Ax (n=1) and Am (n-1). Among 3935 ‘AB’ donors A1B, A2B and weak AB subgroup were 3340 (84.9%), 594 (15.1%) and 1 (0.025%, AmB)) respectively. Anti-A1 (4oC>22oC) with titre ranging from 1 to 8 was found in 127 (21.4%) A2B donors.

Conclusion: Detailed serological characterization of subgroups of ‘A’ and ‘AB’ helps to estimate their frequency in a particular demography. Often weak subgroups may be mistyped as ‘O’ group which may lead to wrong blood transfusion. Often such serological evaluation is needed in solid organ transplantation. Multi-centric studies with large sample sizes are required to estimate the correct frequency of weak subgroups.

2019 May; 13(Suppl 1): S32–S100.

PP 93: A case of passenger lymphocyte syndrome following minor ABO incompatible renal transplantation

Background: Passenger Lymphocyte Syndrome is a rare but important disease in which donor lymphocytes produce antibodies to red blood cell antigens of the recipient causing alloimmune haemolysis. It occurs in ABO blood group mismatched solid organ and/or bone marrow transplant.

Case Presentation: We report a case of passenger lymphocyte syndrome occurring after ABO incompatible kidney transplant. A 45-year-old lady (A +ve) underwent a renal with her husband (O +ve) as donor. The patient was transfused with 5 units of A +ve packed cells (3 units during surgery and 2 units postoperative. The postoperative course was uneventful. However, 15 days postop showed a declining haemoglobin level from 8.6 to 6.1 gm/dl. The patient's sample was sent to blood bank for 1unit of PRBC. The patient antibody screen & DCT were negative. The patient was transfused with 1unit of crossmatch compatible A +ve packed. The patient was discharged following the transfusion. The patient was readmitted 2 days later with a steep fall of Hb to 3.2 gm/dl over 48 hours. There was no identifiable bleeding source. Reticulocyte count ↑ LDH ↑. No shistocytes were observed on the peripheral smear. A request for 2 units of packed cells was again sent to blood bank. However, this time crossmatch with A +ve packed cells showed incompatibility in AHG phase. The antibody screen was negative and DCT was positive (2+). The patient's serum showed presence of anti A. a diagnosis of probable PLS was made. The patient was managed with methylprednisolone & group O PRBC. Gradually her condition improved and was discharged in stable condition.

Conclusion: PLS is an important complication that occurs in patient receiving ABO mismatched transplant. Early diagnosis & appropriate treatment are important in at risk individuals.

2019 May; 13(Suppl 1): S32–S100.

PP 94: Prevalence of abnormal red cell antibody in blood donors: Experience of a North Indian blood bank

Background: The prevalence of antibody screening against the donor's red cell antigen varies from 0.32 to 2.4 % in the literature. The potential hazard of this antibody includes red cell hemolysis of the patients while transfusing plasma and platelets unit. However, red cells are not unaffected in this situation, if SAGM blood bags are used.

Aim: The aim of the study was to evaluate the prevalence of irregular red cell antibody in blood donors at our centre.

Methods: This was retrospective collected data of blood donors at our centre from January 2013 to May 2018. The data of the donors tested for red cell antibody by Indirect Antiglobulin Test (IAT) by erythrocyte magnetic technology was evaluated.

Results: Out of the 56024 blood donation, only 83 samples came as IAT positive. Total positivity was 0.17%. Out of 83 donors, male donors were 67 (80.7%) and female were 16 (19.3%). The percentage positivity in female donors (0.44%) was higher than male donors (0.15%). The majority of IAT positive donors belong to the age group 18-30 years (57 %). Out of the 83 blood donors, 5 were Rh D negative blood group and all of them were female donors. The replacement donors showed higher positivity (0.40%) as compared to voluntary donors (0.12%).

Conclusion: The data showed that out of 674 blood donation, only 1 donor came as IAT positive that showed lesser positivity at our centre as compared to other studies. Higher positivity of IAT in case of female donors and replacement donors has been observed in this study. Minor cross-matching in selected cases like pediatric cases while transfusing FFP and Platelets may be more cost-effective measure as well as less labour intensive in place of donor antibody screening seeing the lesser positivity and risk of transfusion reaction.

2019 May; 13(Suppl 1): S32–S100.

PP 95: Significance of anti-M antibody in patients and donors

Background: Anti-M is a naturally occurring IgM antibody presenting as cold agglutinin. Its conversion to IgG is rare and is associated with hemolytic transfusion reactions and hemolytic disease of fetus and newborn.

Aim: To study the frequency and serological characteristics of Anti-M antibody in patient and donor population.

Methods: It was a retrospective study over for a period of two years (April 2016 to March 2018). Blood grouping was done using conventional tube technique using commercial antisera (anti-A, anti-B, anti-AB and anti-D) and pooled reagent red cells, whereasin case of incompatible crossmatch, antibody screening and identificationwere performed using LISS Coombs’anti-human globulin (AHG) gel cards (Bio-rad, Switzerland). Di-thiothreitol (DTT) treatment was also performed.

Results: The frequency of anti-M antibody was 0.05% in patients (51 out of 101364 requisitions). DTT treatment could be done in 12 patients which showed it was IgM type in 11 (91.7%) and mixture of IgM and IgG type in 1 (8.3%) case. Transfusion history was present in 21 (41.2%), absent in 13 (25.5%) and not known in 17 (33.3%) patients. M antigen phenotyping in 42 patients showed that 35 (83.3%) were M-, while 6 (14.3%) gave a ‘mixed field’ and 1 was M+ as they had a history of recent transfusion. In blood donors, the frequency of anti-M was 0.012% (15 out of 117185 donors) which is lower than the patient population and the difference is statistically significant (p<0.001). This difference shows that the anti-M is not only naturally occurring but may also develop as a clinically significant alloantibody also. On DTT treatment of 9 samples, 4 (44.4%) each were IgM and IgM plus IgG type and 1 (11.1%) was IgG type.

Conclusion: Most of the Anti-M antibodies are IgM type, however, serological characterization reveals the IgG type also which are reactive in AHG phase and thus clinically significant.

2019 May; 13(Suppl 1): S32–S100.

PP 96: Defining serological specificities of multiple antibodies via molecular means: A case study

Background: Allo-immunized patients with multiple blood transfusions pose enormous challenges not only in finding compatible blood but also in antibody identification.

Aim: This study documents our approach in identification of the complex antibodies formed in a multi-transfused patient with sickle cell disease and an adverse obstetric outcome.

Methods: Reagents used for serology work were from local and commercial sources. DNA for molecular typing was extracted from a buffy coat specimen and used for genotyping by Precise type HEA BeadChip (BioArray Solutions, Immucor).

Results: 32 year old 3rd gravida with SCD was admitted to hospital for delivery with Hb of 6.8 g/dL and blood transfusion was planned. She had history of transfusions with episodes of transfusion reactions. Blood group was A1 positive with discrepant reverse typing. Auto-control and the DAT were negative. Three and 11 cell panels showed variable strength of reactivity with her serum which indicated the presence of multiple antibodies to the antigens E, c, S, Fya or Fyb, K, M. Four sequential absorptions with papain treated E-c+ RBCs removed anti-c from the serum leaving other antibodies to identify. Her specimen (plasma) was referred to reference laboratory abroad where her predicted phenotype was determined by molecular testing. Her plasma was tested with selected cell panels as guided by the results by molecular typing. Her predicted phenotype was E-, c-, K-, Fy (a-), S-. Based on testing with phenotypically similar cells, the serology confirmed the antibodies present in her plasma. Only available example of E+, e-, c-, Cw-, Fy (a-) and S- RBCs was reactive with the serum.

Conclusion: Anti-c, anti-Cw, anti-K, anti-Fya, anti-S and possibly anti-E were identified in the patient guided by extensive red cell genotyping data.

2019 May; 13(Suppl 1): S32–S100.

PP 97: Rh-D blocking phenomenon in a case of Rh-D haemolytic disease of newborn

Background: Rh-D blood group system is the most important blood group system other than ABO. Coating of red cells with IgG anti-D antibodies, commonly termed as blocked D, makes the accurate Rh testing difficult.

Case Report: Here we present a case report of Rh-D blocking phenomenon where the blood group of a newborn baby born to a multigravida 32 years old female showed B negataive with the mother's blood group being B negative. The antibody identification of the mother confirmed the presence of allo anti-D along with anti-C. The DCT performed with baby sample showed strong positive (3+) reaction. Antibody identification performed with baby's sample detected the presence of anti-D along with anti-C. Comparison of the results of grouping and antibody detection between the mother and newborn baby led to the elution study performed with baby's DAT positive red cells using cold acid elution method. After elution, baby's red cells were typed as B positive by gel card method.

Conclusion: DAT usually gives strong positive results in HDN due to anti-D alloantibodies. The blocking phenomenon should be suspected if the infant's red cells show strong positive DCT but do not give positive results with anti-D. Before reporting the Negative grouping of a newborn baby of an alloimmunised mother having a good titre of antibodies, especially anti-D antibodies, the Blocking phenomenon should always be suspected and accordingly steps like elution should be done to confirm the grouping.

2019 May; 13(Suppl 1): S32–S100.

PP 98: Evaluation of direct antiglobulin test negative autoimmune haemolytic anaemia: An insight beyond conventional test

Background: A negative DAT does not rule out AIHA. The incidence of this clinical entity known as DAT negative AIHA has been reported to be 2 - 4%. Sensitive techniques like enzyme linked antiglobulin test (ELAT), flow cytometry (FC), complement-fixation antibody consumption test etc. have been described for diagnosis of DAT negative AIHA. Majority of blood bank laboratories lack these advanced methods. Here we share our experience of diagnosing DAT negative AIHA using simple but sensitive methods which are otherwise less practised.

Methods: The prospective study included 377 anemic patients clinically suspected of AIHA. Blood samples received in blood bank were subjected to polyspecific DAT using both conventional tube test (CTT) and column agglutination technique (CAT). Polyspecific DAT negative results were further evaluated using sensitive but simple methods. Hematological and biochemical parameters of all patients were obtained from hospital information system. In vivo hemolysis was categorized as per the criteria established by previous workers. SPSS statistical software (version 13, USA) was used for all statistical evaluation.

Results: Of the final 353 clinical AIHA patients evidence of autoimmunization by CTT was observed in 335. Where DAT negative AIHA was observed in 18 (5.1%) patients, 14 showed evidences of autoimmunization by extended sensitive methods. Four patients responded well to AIHA therapy despite DAT negativity by available methods. Severe hemolysis was observed in 4 (22.2%) DAT negative AIHA patients.

Conclusion: We conclude that DAT negative patients with clinical suspicion of AIHA and positive laboratory evidences should be evaluated for the presence of autoantibody by alternate sensitive methods which are otherwise less practiced. Blood banks may establish these useful simple techniques and stick to the defined protocols to diagnose DAT negative AIHA.

2019 May; 13(Suppl 1): S32–S100.

PP 99: Case report on lewis antibodies detected during routine screening in a tertiary care center

Background: Anti-Lewis antibodies, which are usually not reactive at 37°C, are mostly clinically insignificant. Reports of clinically significant anti-Lewis antibodies causing hemolytic transfusion reactions (HTR) are available.

Methods: Among 6 cases, two individuals were healthy voluntary blood donors, two were patients admitted in our Institute for surgical procedures and two referred cases from local hospital for antibody identification. Routine irregular antibody screening in donated blood units are mandatory as per DGHS guidelines and followed in our Institute.

Results: We report 6 cases of anti-Lewis antibodies reactive at room temperature (RT) and at 37°C. These were found in patients of varied age groups and gender (21 to 65 years) with varied clinical diagnosis. Two pregnant female patients in their second trimester had anti-Le (a) and anti-Le (b) respectively with wide thermal amplitude. Two male healthy voluntary blood donors also were found to be Le (a−b−) with anti-Le (ab) also had wide thermal amplitude. Two male patients with anti-Le (ab) as well as anti-Le (b) respectively with wide thermal amplitude were found during antibody identification after evidence of crossmatch incompatibility. Antigen negative RBC units were transfused in this situation.

Conclusion: Hemodynamic changes in pregnancy may affect certain red cell antigen expression leading to production of naturally occurring antibodies. In case of Lewis the chances of HDFN is extremely rare since Lewis antigens are poorly expressed by fetal cells. Lewis antibodies are usually naturally occurring antibodies present in normal human sera and routine irregular antibody screening in blood donors helps in avoiding untoward events among recipients. Provision of red blood cell antigen phenotyped donor registry shall ensure quick provision of antigen-negative blood for transfusion in emergency situations.

2019 May; 13(Suppl 1): S32–S100.

PP 100: Estimation of anti-A and anti-B antibodies titres in ‘O’ blood group donors for safe transfusion practices

Background: High titres of anti A, anti B antibodies in O blood group donors can lead to potential agglutination of erythrocytes of non-O recipients.

Aim: To estimate risk of dangerous universal donors in blood bank of our state by determining their anti-A - anti-B titres and to evaluate any association with dietary pattern.

Methods: It was a prospective study, conducted during period of December 2017 to May 2018 in SMGS hospital, Government medical college Jammu and consisted of 135 ‘O’ blood group donors. Donors were grouped according to their gender, age and dietary habits. All titration were conducted using tube titration technique. Dangerous donors were those, whose Anti-A or Anti-B IgM titres were greater than or equal to 64.

Results: Among 135 donors (118) 87.8% were male and (17) 12.2% were female. Out of (104) 77% donors who were consuming Mixed diet, (28) 26.9% were dangerous and in remaining (31) 23% donors who were consuming strict vegetarian diet (8) 25.8% were dangerous. In aggregate one third 28.1% (38) of the O blood group donors were universal dangerous. There was no significant association between dangerous universal donors and their dietary habits (p value = 0.932) (non significant <0.05).

Conclusion: Substantial portion of universal donor group, not related to their dietary preferences carry a indigenous risk of dangerous titres against A and B antigen. We recommend prior titration of Anti A and Anti B antibodies of O blood group donors in case of transfusion to recipients of non-isogroup so as to avoid Anti A/B related transfusion reaction.

2019 May; 13(Suppl 1): S32–S100.

PP 101: Management of platelet refrectoriness in a patient of aplastic anemia

Background: Platelet transfusion refractoriness is the failure to achieve the desired level of blood platelets in a patient following a platelet transfusion. The cause of refractoriness may be either immune or nonimmune. Among immune-related refractoriness, antibodies against HLA antigens are the primary cause. Non-immune causes include splenomegaly, fever, and recent chemotherapy. Immune-mediated refractoriness usually shows little or no increment in the immediate post-transfusion platelet count. Non-immune refractoriness may show an initial rise in platelet count, but a subsequent 8 or 12 hour post-transfusion sample shows a return to the baseline platelet count.

Aim: Management of platelet refractoriness in a patient of aplastic anemia.

Methods: The patient, M/26Y, having aplastic anemia got admitted. Hb 6 gm%, platelet count 4000/cumm on admission. Had been transfused more than 100 units of different blood components in last 6 years. He had to undergo major surgery. Received the request of 4 RCC and 8 SDP. After transfusion of 2 RCC, Hb improved but even after transfusion of 6 SDP, platelet count remained static 4000/cumm, without showing a bump up at any point of time. Two more SDP, but no improvement. As there was no bump up in platelet count for a brief period even, we suspected immune-mediated platelet refractoriness. To overcome this, we had the only option of PLATELET CROSS-MATCH. It was carried out by SPRCA (CAPTURE-P), NEO, IMMUCOR.

Results: Blood group “A” Positive. DAT, IAT and AUTOCONTROL negative. Total 12 potential SDP donor's samples were cross-matched with patient sample. 4 of those gave best compatibility. After 1st SDP transfusion, platelet count increased from 4000 to 20000/cumm. Then 2nd SDP given and platelet count reached to 50000/cumm. This way patient not only came out of impending bleeding crisis but was operated successfully.

Conclusion: Platelet crossmatch is a boon to manage platelet refractoriness in a speedy and effective way.

2019 May; 13(Suppl 1): S32–S100.

PP 102: Occurrence of clinically significant lewis antibodies (Anti-lea) in voluntary blood donors - Two case reports

Background: Lewis antibodies are usually naturally occurring and of the IgM class. Unless acquired by sensitization from previous transfusion, they are not clinically significant. It is even rare to find idiopathic presence of anti-Lea and anti-Leb which can cause adverse reactions. Sensitive assays are required to detect these IgG type of Lewis antibodies.

Aims: To share our experience from two cases with presence of clinically significant IgG class of Lewis antibodies among voluntary whole blood donors for a tertiary care oncology centre in Mumbai.

Methods: The blood samples of consenting blood donors were tested routinely as per institutional Standard Operating Procedures (SOPs). Blood grouping and antibody screening were done using commercially available antisera by column agglutination technique (CAT) and ‘O’ cells. Reactivity in both samples necessitated further analysis by 3-cell and 11-cell antibody screening panel. The samples were tested at room temperature (RT), 370C and Indirect Antiglobin test (IAT) phases.

Results: The respective blood groups being ‘O’ Rh positive and ‘B’ Rh positive in routine blood typing, both samples showed reactivity at 370C and Indirect Antiglobin test (IAT) phase on 3-cell panel and 11-cell panel which delineated Anti-Lea in both. This indicated that the antibodies were probably of IgG type. There was no significant history of sensitizing factors in both donors.

Conclusion: These two cases serve to highlight the presence of Lewis antibodies among donor population which are generally of IgM type. Anti-Lea is the most commonly encountered of the Lewis antibodies and is often detected at RT, 37°C and in IAT phases. Incorporation of testing with potent commercially available antibody specific antisera and extended blood group phenotyping should thus be considered.

2019 May; 13(Suppl 1): S32–S100.

PP 103: Advances in transfusion technics is a boon for intrauterine transfusion - A case study

Background: Foetal anaemia has many causes. Haemolytic disease secondary to maternal (Rh) isoimmunisation is the commonest cause, despite prophylaxis of anti-Rh imunoglobin. Approximately 10% of isoimmunised women have a child affected in the uterus by haemolytic anaemia. Proper pregnancy monitoring enables correct time for the treatment of anaemia and foetal hydrops. Hence new methods of diagnosing and treating this disease are necessary. Before ultrasound technology, the foetal effects of Rh isoimmunisation were detected only at birth. Sensitised Rh-negative pregnant women are monitored using signs of foetal haemolysis & anaemia corrected through intrauterine transfusions (IUT).

Methods: A 29 yrs Fifth gravida with bad obstetric history is being treated by Obstetrician for antenatal check up. Considering her history blood group, Anti D titer, Cell panel study was done at 20th wk. Blood group discrepancy was found (Cell A & serum O Rh negative), Anti D titer – 1: 1024, Anti D & anti C antibodies were found. Meanwhile foetal anaemia was noted by assessing the peak systolic velocity in the foetal middle cerebral artery (PSV-MCA). The first IUT was performed when hemoglobin concentration was less than 5 g/dL & haematocrit <30%. O Rh Negative fresh PCV (40 ml), with log 4 leuco- reduction, NAT tested & Irradiated unit was transfused. Baby's Hb transiently raised & again dropped. So second IUT was given with same specifications after 48 hrs. Baby's Hb had raised & baby is active in womb.

Results: With expert hands of Obstetric team & supply of required blood components with required specifications, baby showed normal intrauterine activities on ultrasound examination, monitering wkly upto 34 wks.

Conclusion: For patients with bad obstetric history one need to have prompt team efforts & transfusion support in time for a birth of healthy baby, indicates the authenticity of advanced transfusion technologies.

2019 May; 13(Suppl 1): S32–S100.

PP 104: A rare combination: X-ray irradiation and Bombay blood group

Background: Irradiated blood products are required to prevent TA-GVHD. Patients who are immune-compromised and receiving transfusion from a relative (directed donation) are at increased risk of it. Typically it occurs 10-14 days of post transfusion and is fatal in more than 90% of cases. Bombay blood group is a rare group with the incidence of about 4 in 10,00,000 in India, in which there is absence of H antigen and presence of anti-H antibodies. So, at time of blood grouping, it looks like O group due to absence of H antigen. Thus, they can donate blood to anybody but can receive blood only from same group people.

Aim: To transfuse a rare group from a relative (directed donation) in the safest possible manner.

Methods: We at LBC, received blood samples of M/45 Yrs, requesting for 2 RCC for transfusion, admitted for surgical procedure. As routine practice, both forward and reverse blood grouping was carried out using Tube Technique. Simultaneously, antibody screening was done using Gel Technology. We found a discrepancy in blood grouping so patient's RBC was tested against anti-H lectin, which confirmed O Bombay. Patient's siblings were screened. X-ray Irradiation came into picture to prevent TA-GVHD.

Results: Forward group was O positive while in reverse, there was an unusual reaction with O cells. Reaction with anti-H lectin was negative which suggested the absence of H-antigen. In IAT the reaction was 4+ with each cell. Patient's two brothers were identified as having Bombay phenotype. Both were undergone through donation criteria and successful donation done. Compatibility testing done by CAT method and RCC was issued after X-RAY Irradiation process to avoid TA-GVHD as the donation was from a sibling.

Conclusion: “X-RAY Irradiation” made it safest to transfuse a rare “Bombay group.”

2019 May; 13(Suppl 1): S32–S100.

PP 105: Rare case report of neonatal hyperbilirubinemia in twins due to Rh hemolytic disease of fetus and newborn

Background: Haemolytic disease of the newborn (HDFN) is a very common condition that leads to significant hyperbilirubinemia and anemia in the neonatal period. This condition is secondary to maternal antibodies crossing the placenta and producing hemolysis by destroying the fetal or neonatal red blood cells (RBC) leading to hyperbilirubinemia in neonatal period often requiring phototherapy, exchange transfusion or blood transfusion later in life. Here we report a case in which both the twin had severe hyperbilirubinemia due to anti ‘c’ antibodies.

Case Presentation: Samples of mother (P2L3) who recently delivered twins were sent to us from nearby local hospital with history of neonatal jaundice in both twins. On grouping both mother and neonates were O positive. DAT was positive for both neonatal samples and maternal sample showed a positive IAT. The irregular antibody present on the maternal serum was identified as anti-c with a titre of 128. On Rh antigen phenotyping, mother was CCDEe. ZZAP treatment of both neonatal samples was performed till DAT came negative. The antigen Rh phenotyping of twins revealed to be CcDEe. The father and first child were also O positive and Rh phenotyping gave ccDEe and CcDee respectively. There was no previous history of blood transfusion for mother. The mother was sensitised in her first pregnancy that lead to anti-c production which affected her subsequent pregnancy. Both neonates had immediate post partum serum bilirubin values 17.1 and 17.3 mg/dl and hemoglobin values 13.5 and 13.9 g/dl respectively who required phototherapy on Day1. 24 hr post phototherapy serum bilirubin values were 13.2 and 13.7 mg/dl and hemoglobin values were 12.7 and 12.1 g/dl respectively and exchange transfusion was not required for them. Both mother and infants were discharged on Day 5 with no further complications.

2019 May; 13(Suppl 1): S32–S100.

PP 106: Bystander hemolysis in sickle cell disease: A case report in siblings

Background: Alloimmunization in sickle cell disease (SCD) may cause delayed hemolytic transfusion reaction/hyperhemolysis (DHTR/H) syndrome. In some instances of DHTRs, the apparent loss of circulating red cells exceeds what would be expected if only antigen-positive transfused red cells were cleared from the peripheral blood. There may be evidence of complement deposition on autologous red cells, with a positive direct antiglobulin test (DAT) due to C3 persisting for up to 100 days. This phenomenon has been called “bystander hemolysis”. Typically, bystander hemolysis is mild and is differentiated from autoimmune hemolytic anemia (AIHA) by the absence of anti-IgG reactivity in the DAT and the lack of autoantibody in the eluate.

Aim: We report cases of brother and sister (aged 14 and 13 respectively) of SCD presenting with pain crisis. They had signs and symptoms of accelerated hemolysis evidenced by an unexplained fall in Hb, elevated Lactate Dehydrogenase, elevated bilirubin, and hemoglobinuria, all occurring between 7 to10 days after RBC transfusion.

Methods: ABO grouping and auto-control (4 degrees, 37 degrees and room temperature) were done by conventional tube technique (CTT). DAT, antibody screening, antibody identification were done by column agglutination technology (CAT) using orthoBioVue card and Ortho reagent red blood cells (surgiscreen and Resolve PanelA).

Results: Both had DAT positive (only C3d with IgG negative). Antibody identification of both showed anti-E alloimmunization with negative auto-control. They were successfully managed with avoidance of further transfusions and administration of corticosteroids.

Conclusions: DHTR/H syndrome in SCD patients may mimic pain crisis. HTRs may be severe in patients with SCD. In such patients, the degree of anemia may be greater than before transfusion. This is most likely the result of bystander hemolysis of autologous red cells. Pain crisis in SCD patient following transfusion, or the inability to sustain an adequate hematocrit, should suggest the occurrence of sickle cell HTR syndrome. It is important to realize that further transfusion in this setting may exacerbate the anemia and even prove fatal.

2019 May; 13(Suppl 1): S32–S100.

PP 107: Blocked - D phenomenon in a newborn with passively acquired maternal anti-D and anti-C

Case Report: A 27-year-old lady with one live birth and one abortion was admitted at 35 weeks of gestation. Her blood group was A-Rh (D) negative and first child A-Rh (D) positive. She received Anti-Rh (D) prophylaxis after delivery and abortion. No history of blood transfusions. Indirect Antiglobulin Test (IAT) was positive with a titre of 64 at 16 weeks of gestation. From 24 weeks, fetal anemia warranted intrauterine transfusions. Last transfusion was given five days prior to delivery. Baby was delivered at 35 weeks of gestation. Baby's blood group on cord blood sample was O-Rh (D) negative. Being a case of fetal anemia requiring intrauterine transfusions, cord blood samples of baby and maternal blood samples were sent to rule out Hemolytic Disease of Fetus and Newborn (HDFN). Blood group of mother was confirmed as A-Rh (D) negative. IAT was positive with a titre of 512. Antibody screening and identification showed pattern corresponding to “anti-D” and “anti-C”. In order to rule out anti-G, maternal serum was adsorbed onto R2R2 (D+C-) cells. Eluate from these cells tested for reactivity against r’r (D-C+) cells were negative. Hence, anti-G was ruled out. Maternal Rh phenotype was D-C-E-c+e+. The antibody titre against R2R2 (D+C–) cells were 512 and r’r (D–C+) cells were 32. Thus, anti-D titre was 512 and anti-C was 32. Cord blood sample of baby was initially typed as O-Rh (D) negative. Direct anti-globulin test showed weak mixed field reaction. Suspecting blocked-D phenomenon, baby's RBC subjected to elution showed weak mixed field reaction with anti-D revealing baby to be Rh (D) positive. Weak reaction could be explained by the presence of RBC from recent intrauterine transfusions. C antigen typing was negative. Eluate showed reactivity pattern of anti-D.

Conclusion: This was a case of HDFN presenting as blocked-D phenomenon due to transplacentally obtained anti-D with coexisting anti-C.

2019 May; 13(Suppl 1): S32–S100.

PP 108: Pretransfusion antibody identification: A retrospective analysis of a tertiary healthcare system

Background and Aim: The objective of pre-transfusion antibody screening for red cells is to ensure that enough red blood cells (RBCs) survive when transfused. This retrospective study was conducted with the aim to see the prevalence of clinically significant alloantibody/autoantibody.

Methods: This study was carried out in a tertiary healthcare center between June 2015 and May 2018. There were a total of 19888 blood samples of patients tested for 3 cell antibody screening by solid phase method using Immucor Capture-R Ready Screen 3. Positive antibody screen were further tested for antibody identification using Immucor Capture-R Ready-ID Panels. In case of presence of autoantibody, presence of alloantibody was detected using Immucor's Elukit & WARM reagents. Freshly prepared cord blood was used, when required. Multiple panels and additional reagents were only used for any unresolved or in autoantibody cases.

Results: Of 177 positive screen samples 111 (63%) sample were identified for having single or multiple allo- antibodies followed by 53 (29%) samples having auto antibodies with AIHA and 13 (7%) results were found inconclusive having nonspecific antibodies. Of allo-antibodies, total 129 antibodies were identified in single or multiple doses, out of them 111 (86%) of total identified antibodies were Rh & Kell type (anti-D-52%, anti-E-16.2%, anti-c-7.7%, anti-C-4.6%, and anti-K-4.6%). In total 53 AIHA cases 43 (81%) samples were tested having only warm reacting auto-antibodies and 10 (19%) were having autoantibody along with other allo-antibodies. Other clinically significant antibodies detected along with auto-antibodies were anti-Fyb, anti-Jka, anti-Jkb, anti-S, anti-P. In 1 case (2%) autoantibody was tested with specificity of ‘Anti-D’. And 2 (4%) samples were identified as Cold AIHA with specificity of ‘anti-I’.

Conclusion: The alloantibody detected were mainly against the Rh and Kell phenotype (>85%). Approximately 20% of the AIHA cases were having hidden alloantibodies.

2019 May; 13(Suppl 1): S32–S100.

PP 109: The double edged sword of heightened sensitivity - tubes, columns and titers!!!

Background: The time tested method of doing antibody titration is by tube method. With the advent of Column agglutination technology and the ability to automate it is tempting to assess the comparability of this platform in determining titers.

Methods: This study was done with the aim of comparing Coombs titer of 5 patients and 4 different lots of antisera on tube and column agglutination technology method. For the latter, testing was done on Ortho and Biorad cards.

Results: Of the reagent titers tested on all three methods, despite difference in titers it met quality control requirements. The difference noted varied as follows. Compared to tube technique, Ortho column showed higher titer equivalent to one tube. However, Biorad showed one tube difference for one lot and two tube difference for three reagents. With regard to patient titers, the titers of 2 and 4 correlated well with the Biorad column, but showed marginally increased titer equivalent to two tube difference in the Ortho card. Titers of 8 showed and one and two tube difference and 16 showed a two and three tube difference in Ortho and Biorad respectively.

Conclusion: Column Agglutination Technology is clearly a more sensitive platform and higher titers are to be expected if performed using the same. In the case of reagents, the impact on working processes was minimal as all of them were clearly within acceptable quality control ranges. However, in patient samples particularly, in titers of 8 and 16, the discrepancy noted would lead to changes in monitoring and management. It is therefore important to document clinical correlation of titers identified on the Column Agglutination platform before they can be used for clinical care. The difference between various vendors requires attention as well.

2019 May; 13(Suppl 1): S32–S100.

PP 110: Prevalence of irregular red blood cell antibodies among healthy blood donors in South India

Background: Although antibody screening of donors is mandatory according to National blood policy of India, the prevalence of irregular red cell antibody and its specificities are not much reported in South Indian population.

Aims: To find the prevalence of irregular antibodies and positive Direct antiglobulin test (DAT) in healthy blood donors in South Indian population.

Methods: This is a retrospective observational study conducted in the department of Transfusion Medicine from April 2011 to March 2018 (7 years) at a tertiary care referral centre, Chennai, Tamil Nadu. Red cell antibody screening of all voluntary blood donors was performed using Commercial O cells (ID Diapool, Diamed Cressiersur Morat, Switzerland). Positive sera were further investigated to identify the irregular erythrocyte antibody by commercially available red cell panel (ID-Dia Panel, Diamed-ID Microtyping System).

Results: A total of 40,629 donors were screened for the presence of irregular erythrocyte antibodies. A total of 57 donors showed presence of irregular antibodies in which 41 (71.9%) were alloantibodies. Most frequent alloantibodies identified were of MNS blood group system with anti-M being highest (n=15, 36.58%) followed by Rh Blood group system 17% (n=7). 16 donors were DAT positive which were detected during routine cross matching. Prevalence of antibodies were higher in females (0.5%) compared to males (0.12%). Majority of the alloimmunized donors were in the age group of 25-44 years.

Conclusion: Implementation of red cell antibody screening in all the blood donors routinely helped us understand the prevalence of antibodies in our region and its importance in providing compatible blood products and to avoid transfusion reactions.

2019 May; 13(Suppl 1): S32–S100.

PP 111: Differentiation of anti-D + C from anti-G by differential adsorption and elution method in a resource constrain setup

Background: The G antigen is found on red cells possessing C or D antigen. Anti-G antibody serologically mimics a combination of anti-C and anti D antibodies. The challenge of anti G in the antenatal setting is to identify whether anti-D is present or not. If anti-D is absent, the female can still get immunized against D antigen and it warrants RhIG prophylactic therapy. Hence differentiating them is important as anti-D + anti-C cause's severe haemolytic disease of the foetus and new born than anti-G.

Aims: We have differentiated anti-G from anti-D+C by differential adsorption and elution method to determine the need of RhIG prophylaxis.

Methods: Sera from antenatal mothers, whose antibody identification by 11-cell panel (Dia Panel, Bio-Rad, Switzerland and Ortho clinical Diagnostic) gave a pattern for anti-D and anti-C were selected. The serum of antenatal women were adsorbed by differential adsorption at 370C for 1 hr and elution was done by gentle heat elution (at 450C) using R2R2 cells and r’r cells to distinguish anit-G from anti-D + anti-C. Extended phenotyping (conventional tube technique) for Rh system was performed for these antenatal women. Antibody titres of these antibodies were determined and their clinical outcome in the new-borns were followed.

Results: A pattern suggestive of anti D and anti C combination were observed in all the four antenatal cases. After adsorption and elution all of them confirmed to have anti D and anti-C combination. Antibody titres of anti-C were lower than that of Anti-D. All new-borns were sensitized in vivo and the antibody specificity in them were confirmed with elution studies. All the mothers who had anti-D+C so they didn’t need RhIG prophylaxis.

Conclusion: Differential adsorption and elution studies help in distinguishing anti-D plus anti-C from anti-G, thus helping in better patient management.

2019 May; 13(Suppl 1): S32–S100.

PP 112: Rh antigens and phenotypes in a population of blood donors at a tertiary care medical institute of rohilkhand region, Uttar Pradesh, India

Background: The Rh blood group system is most immunogenic next to the ABO system. Allo-immunization remains a risk for individuals who are transfused with red cells carrying antigens that are deficient on the recipient's red cells. It is imperative for blood transfusion services to be aware of the red cell Rh antigen types and their phenotypic expressions in their local and regional blood donor population.

Aims: The present study was therefore undertaken to observe the frequency distribution of common Rh antigens C,c, E, e, and various phenotypes among blood donors; and further, to evaluate their possible association with gender and major blood groups (A, B, O, AB and Rh- D).

Methods: This study enrolled 9091 blood donors. Antigen typing was done by fully- automated immuno- hematology analyzer (NEO) using monoclonal antisera (Immucor, USA). Frequency distribution of antigen types and various phenotypes was observed. Chi-Square test was applied to evaluate their possible association with respect to gender, Rh- D status and ABO type, using statistical software SPSS version 23. Results were considered statistically significant at 95% significance level.

Results: In the present study, e was the most common antigen with a total prevalence of 98.86%, followed by C (85.05%), c (55.03%) and E (18.03%). Prevalence of D was 94.37%. CCee (44.69%) was the most common phenotype, while CCEe (0.27%) was least common. Other phenotypes included Ccee (28.93%), CcEe (11.11%), ccee (8.34%), ccEe (5.50%) and ccEE (1.14%). No statistically significant association was observed for phenotypes with respect to gender, Rh- D status and ABO blood group; however the result was statistically significant when antigen frequencies were compared with blood groups of ABO system (p<0.00001).

Conclusion: A varied frequency of Rh antigens and phenotypic expressions in blood donors necessitates a more rational approach for blood transfusion with appropriate antigen matched blood, to enhance blood safety.

2019 May; 13(Suppl 1): S32–S100.

PP 113: Role of blood transfusion services in detecting para neoplastic autoimmune hemolytic anemia in uterine cancer

Background: Autoimmune Hemolytic Anemia (AIHA) is a paraneoplastic process associated with lymphoproliferative disorders and rarely associated with solid tumors. It may be associated with immune platelet antibodies (evans syndrome), lupus anticoagulants or antibodies to C1esterase inhibitor. AIHA is associated with tumors arising from almost all organs as lungs, breast, prostate, urinarybladder, kidney, uterus, pancreas, thyroid, liver, ovary, stomach, cervix, testis. AIHA associated with the tumors is mediated by warm &/cold antibodies.

Case Report: A 58 year old postmenopausal lady diagnosed with endometrial adenocarcinoma grade II underwent staging laparotomy in 2013. She defaulted for 4 years and presented with bleeding per vaginum for 10 days. Ultrasound indicated hypoechoic mass 1.5*1.4 cms in vault. Biopsy indicated poorly differentiated adenocarcinoma (grade III) and was diagnosed as vault recurrence. Chemotherapy with Taxol/Carboplatin was given for 6 cycles. She was assessed for radiotherapy. Mean while, patient was advised blood transfusion for low hemoglobin. There was difficulty in crossmatch and fresh blood sample was requested for Coombs/Antiglobulin test. Direct and Indirect Coombs test and Autocontrol were positive. The laboratoty findings were also suggestive of AIHA. Two units of Least Incompatible PRBC were issued and Hemoglobin increased.

Conclusion: It is identified that AIHA may occur prior to, concurrent with cancer or well after end of treatment,either as a sign of recurrence or in complete remission of the cancer. Tumors of breast, lung, renal cell, bladder, ovary and cervix have been reported where AIHA occurred at the time of recurrence and was not present at initial diagnosis of cancer. In this case of uterine cancer, AIHA occured at the time of recurrence. The Oncologists may work in close association with Blood Bankers to diagnose AIHA as a marker of disease activity in solid tumors and try to establish the complex immune mediated pathogenesis associated between Cancers and AIHA.

2019 May; 13(Suppl 1): S32–S100.

PP 114: Prevalence of Bombay phenotype in a tertiary care hospital in Chennai

Background: Bombay blood group (Oh phenotype) first discovered in Bombay, by Dr. Y.M. Bhende in the year of 1952. Individuals with the rare phenotype (hh) do not express H antigen (also called H substances). Bombay phenotype can donate RBCs to any member of the ABO blood group system if the Rhesus (Rh) antigens are compatible. They must receive blood from the people who have the Bombay phenotype only. In the general population the prevalence of Bombay blood group is about 1 in 10,000 individuals in India and 1 per 1,00,000 individuals in Europe1.

Aims: To determine the prevalence of Bombay phenotype in a tertiary care hospital, Chennai.

Methods: The ABO and Rh-D typing were carried out in blood donors & recipients by standard cell and serum grouping by test tube method using commercially available anti sera and known cells prepared in house, from pooled blood units. All blood samples showing “O” group were tested with commercial anti-H lectin of Ulex europaeus seed extract for the period of 5 years in Govt. Royapettah Hospital, Chennai.

Results: Analyzing the results of 31410 study subjects showed that the most common was “O” group (39.6%); 5 oh phenotypes (0.016%) were detected of which 4 were males and 1 female, and all were Rh-D positive.

Conclusion: All blood group “O” blood donors and recipient be routinely screened for Bombay phenotype to reduce the risk of a patient with Bombay phenotype being transfused blood group O blood and causing a fatal blood transfusion reaction. There is need for the implementation of a serum typing or reverse grouping confirmation along with ‘O’ cell control in reverse grouping procedure in blood transfusion laboratories to detect Bombay blood group, which is commonly mistaken as “O” Blood group.

2019 May; 13(Suppl 1): S32–S100.

PP 115: Serological “weak D” phenotype: A double trouble - is genotyping the only solution?

Background: Complexity of the D antigen expression has led to inconsistencies in transfusion medicine. There is variation in reporting of Rh D typing results when testing for Rh D by different methods. Lack of standardized testing and reporting leads to discrepancies when a patient consults at different centres. This issue is pertinent in antenatal cases with regard to administering RhIg.

Aim: To estimate the prevalence of serological weak D among the patient population in a tertiary care hospital.

Methods: Patient blood samples received in the blood bank of Sri Ramachandra Medical College & Research Institute, Chennai for ABO & Rh typing during the period January 2017 to June 2018 were subjected to testing by the conventional tube technique. For Rh “D” testing, IgM anti-D reagent was used. Whenever a sample showed a weaker reaction for Rh “D”, further testing for weak D was carried out using IgG anti-D reagent.

Results: During the study period, 58,800 blood samples were subjected to ABO & Rh typing out of which 124 samples were serological weak D; of these 124 serological weak D, 26 were antenatal cases.

Conclusions: Prevalence of serological weak D was 0.2% in the present study. These individuals are to be considered Rh “D” negative as transfusion recipients. If these serological weak D individuals are proven to be Rh “D” positive by genotyping, precious reserve of Rh D negative donor blood can be spared for the confirmed Rh negative patients in case of transfusion needs. Genotyping for RhD can help in resolving the issue of D variants like weak D and Partial D. Consistent practice is important when reporting the Rh D status for all patients, more so in antenatal cases to avoid unnecessary administration of RhIg.

2019 May; 13(Suppl 1): S32–S100.

PP 116: A rare case of Bombay Rh D negative phenotype in a pregnant patient

Background: Bombay phenotype or Oh is a rare autosomal recessive phenotype characterized by absence of H, A and B antigens on red cells and in secretions. Bombay phenotype is challenging when patient needs transfusion. We report an interesting case of Rh (D) Negative Bombay phenotype in a pregnant woman.

Case Report: A 26-year-old pregnant woman G2P1L1was referred at 28 weeks gestation to the Obstetrics and Gynecology OPD. Patient was typed as O Rh (D) negative with positive indirect antiglobulin test (IAT) elsewhere. On blood grouping, discrepancy between cell and serum grouping was observed (cell grouping showed as O negative and serum grouping gave positive reaction with O pooled cells). IAT of patient was positive with 4+ strength using column agglutination technique. Auto-Control and Direct Antiglobulin Test (DAT) were negative. Patient's sample was further tested with Anti-H lectin along with O cells as control and she was confirmed as Bombay Rh D negative phenotype. Past history revealed transfusion reaction with O negative blood. Family screening revealed Bombay Rh D positive in one of the siblings. Multiple adsorptions were performed with O negative cells to adsorb Anti-H from serum. After adsorption, IAT was negative with adsorbed serum at AHG phase. IgM Anti-H titer was 1:32 and IgG was 1:64. The clinician was informed about the rarity of blood group, difficulty in finding donor of same group, need for Anti D immunoprophylaxis and close fetal monitoring. Her initial hemoglobin was 9.0 g/dl. Dietary advise and hematinics were added to build up her hemoglobin to 11 g/dl. An elective LSCS was planned and she delivered a healthy child at 36 weeks of gestation without any blood transfusion.

Conclusion: The case highlights the importance of grouping with inclusion of O cells in serum grouping and the challenges faced by blood bank to provide rare blood phenotypes.

2019 May; 13(Suppl 1): S32–S100.

PP 117: Ax subgroup: A case report

Background: Weak subgroups of A can be defined as those of group A subjects whose erythrocytes give weaker or no reactions with anti-A antisera than do those of subjects with A2 RBCs. These weak phenotypes, in majority of the cases result from the expression of an alternate weak allele present at the ABO loci. Weak subgroups of A are A3, Ax, Aend, Am, Ay, and Ael.

Methods and Results: Qwalys 3 (Diagast) show discrepancy in cell & serum grouping. Then ABO grouping was done by conventional tube technique after washing the donor and reagent RBCs three times with 0.9% normal saline. Monoclonal anti-A and anti-B and anti-A, B antisera were used to reconfirm the routine findings. Agglutination of donor red cells by anti-A1 and anti-H lectin was also determined. The result was confirmed after microscopic examination. For confirmation of blood group adsorption-elution and saliva testing was done. The results of above tests show that the blood group of donor was AX Rh D positive.

Conclusions: Identification of Ax subgroups is important because these donors may be mistyped as group O. It is very dangerous when these units transfused to O blood group patients. This is due to naturally occurring anti-A and anti-B antibodies present in O blood group patients.. Similarly since Ax individuals almost always have anti-A1 antibodies in their serum. If clinically significant, they can lead to fatal transfusion reactions on transfusing their whole blood or plasma to group A individuals. All the donors should personally inform about their group and issue a special blood group card.

2019 May; 13(Suppl 1): S32–S100.

PP 118: Aint blood group: A case report on rare subtype of blood group A

Background: A1 (80%) and A2 (20%) are the major subgroups of A. This differentiation is based on the basis of reactivity of A1 cells but not A2 cells with anti-A1 lectin. A2 cells show increased reactivity with anti-H lectin. The role of subtyping group A is critically important as A2 organ can be transplanted to O group not A1 group. The incidence of Aint subgroup is unreported in this region.

Case Report: A 23-years male came to Blood Bank, AIIMS Raipur as replacement donor. He was screened and found fit for donation. 350 ml whole blood was collected. All the pre-donation, donation & post-donation protocols were followed. After donation, blood grouping was done by tube method. Forward grouping was done with commercial monoclonal antiseras anti-A, anti-B, anti-D1 and anti-D2 (monoclonal anti-D antiseras from different lots) and reverse grouping done with DAT negative pooled A cell, B cell, O cell and Autocontrol. It came out to be A-positive. It was further tested with anti-A1 lectin which gave 1+ reaction. On further testing with anti-H lectin, it showed 4+ reaction. On saliva testing, both A, and H substances were present. Based on these results it was typed as subgroup of A- A intermediate (Aint) group. However we were not able to perform molecular tests.

Conclusion: Aint is considered a heterogeneous subgroup commonly seen in black people than in white ones. In India, the incidence of Aint was reported to be 2% in Maharashtra. There is no reported case of Aint in this region and this would be the first reported case. Our report points to the need to perform molecular tests or flow cytometric analysis in certain cases. We also recommend that Anti-A1 and anti-H lectin must be mandatorily used for typing of all A blood group.

2019 May; 13(Suppl 1): S32–S100.

PP 119: Blocked D phenomenon: A case report

Background: Blocked D phenomenon is one in which all the D antigens on fetal D Pos cells are coated with maternal IgG Anti-D, making fetal red cell typing difficult. It is encountered in cases of severe HDFN. The D antigen of Rh system being most immunogenic is the most common cause of HDFN. Since the Rh antigens are fully expressed at the time of birth, unlike the weak expression of ABO in neonates, confirming the Rh status can be difficult at times. Here we report a case of Blocked D in a 2 days old infant born to a Rh negative mother.

Case Report: Sample of 2 days old male baby c/o suspected HDFN was received for confirmation of grouping and DCT. Mother was 28 yr old G4P2L2A1 case of Rh incompatibility. Mother's blood grouping was found to be AB Neg and father's group was B Pos. Mother's antibody screen and identification revealed anti-D with a titre of 1:256. Baby had jaundice on day 1 of life with total bilirubin of 19.6 g/dl. DCT was 4+, but the grouping was found to be AB Neg. Hence grouping was repeated with warm saline wash in conventional tube technique. Repeat grouping was found to be the same (AB Neg). Heat elution was done to remove the antibodies coating the red cells. Antibody identification of eluate from the baby sample revealed anti D. Grouping of eluted cells was found to be AB Pos.

Conclusion: Blocked D phenomenon is believed to be far more likely in theory than in practice. With the use of commercial monoclonal reagents, rate of false negative Rh typing is usually very less. However, if not identified promptly, it can cause delay in making a diagnosis Rh-HDFN and further management.

2019 May; 13(Suppl 1): S32–S100.

PP 120: When safety is in numbers - the role of multiple reagents and platforms in Rh (D) typing

Background: Assigning Rh (D) status confidently is extremely important in the practice of transfusion medicine. The immunological implications of wrongly assigned Rh (D) status are well known and have implications of great magnitude. We report here a case of a 12-year-old female child in whom a discrepancy of Rh typing related to the reagent used was observed.

Case Report: A request was received for a unit of RC for a 12 years old female child with Glanzmann Thrombasthenia. As per institutional protocol ABO and Rh typing was performed using both tubes and column agglutination platform (CAT). The former showed a clearly negative reaction with Anti D whereas latter showed 3+ reactivity. The tube was subjected to weak D testing which showed a 3+ reaction. Given this difference the sample was retested using tube technique with alternate reagent and 2 different CAT and positivity noted in all these. The clear negativity noted on the tube technique with just one reagent raised the possibility of this being reagent related. In order to address this, the test was repeated with 4 different lots of the same reagent which were again clearly negative. Titers in all of these lots met quality control requirements. Subsequently molecular typing using PCR SSP was performed for both weak D and partial D. Results showed it to be clear Rh positive.

Conclusion: This D positive child who demonstrated negativity with one particular reagent demonstrates the challenge of assigning D antigen status when there is variability of reagent reactivity. It was strange that 4 different lots of a particular D typing reagent continued showing negativity while 4 other reagents on 2 different platforms showed positivity which was confirmed by molecular typing. This case highlights the need for multiple platforms and multiple reagents for immunohematology testing.

2019 May; 13(Suppl 1): S32–S100.

PP 121: Overcoming daratumumab interference in immunohaematology tests – A case report

Background: Drug interference is a well-known phenomenon in Transfusion medicine. Interference with routine methods for compatibility testing for blood transfusion puts patients at risk for delay in receiving compatible blood. Daratumumab, a novel IgG1k anti-CD38 monoclonal antibody which effectively targets human myeloma cells, shows an unexpected interference in routine immunohaematology tests.

Case Report: 49 year old male, a known case of Relapsed refractory IgG-K multiple myeloma had disease relapse on day +135 post autologous stem cell transplant (SCT). It was planned to get him into remission with novel agents followed by matched sibling donor allogeneic SCT. Despite therapy, he had progressive disease with increasing monoclonal protein, and he was started on monoclonal antibody Daratumumab based protocol. Initial antibody screen using commercial three cell panel, Direct and Indirect AHG were negative. He received 11 units of AHG compatible packed cells pre-Daratumumab therapy. Extended phenotyping could not be done as the previous transfusion was within three months. After the initiation of Daratumumab therapy, antibody screen, direct and indirect AHG became pan reactive (2+). Crossmatch with multiple units were compatible upto 370C, but incompatible at AHG phase. In order to eliminate the interference of anti-CD38, donor units and reagent cells were treated with dithiothreitol (DTT). After DTT treatment with standard protocol the antibody screen was negative and the RBC units were AHG-compatible. The patient was transfused with compatible RBC units and no hemolytic transfusion reactions were observed. Since the RBC units were AHG compatible prior to the initiation of daratumumab, we suspected that daratumumab interfered in pretransfusion testing.

Conclusion: Daratumumab interferes with the antibody screening, causing a panreactivity that can mask clinically significant allo-antibodies. Routine serological methods are ineffective in circumventing the interference in compatibility testing, there by resulting in an unexpected delay in finding suitable blood for these patients.

2019 May; 13(Suppl 1): S32–S100.

PP 122: Does red cell alloimmunization continue to challenge breast fed babies? A unique case report

Background: Nature has provided adoptive immunity in two instalments during early mammalian life. As pregnancy progresses, the maternal immune system shifts away from Th2- and towards Th2-driven responses, a unique humoral bias which facilitates the transmission of maternal IgG across the placenta, while IgA largely in extrauterine life through breastmilk. We present a case report of IgG red cell antibodies being transferred through breastmilk and persisting at six months. Literature search showed this to be the second case being reported worldwide.

Case Report: Blood sample of a six month old child was received for blood grouping and compatibility testing. Antibody screen was positive and alloantibody with anti-Kell specificity was identified. Careful revisiting of the history showed no sensitising events. Hence the mother was tested and was found to have anti Kell antibodies. The maternal IgG in newborn declines progressively over 1-2 weeks as the antibody slowly breaks down. These observations led to the question of whether IgG RBC alloantibodies continued to be passively transferred from mother to child via breastmilk. Therefore consent was obtained and breastmilk was tested which confirmed positive for anti-Kell specificity. The titres of these antibodies in maternal serum, breastmilk and baby's serum were found to be 1:1024, 1:2 and 1:8 respectively. Transmission through breastmilk may be due to the markedly high titers in mother. Monospecific DAT with breastmilk showed IgG isotype. Further, subtyping of IgG showed presence of IgG1 in both maternal and baby's serum but not in breastmilk owing to very low titers. No evident hemolysis was seen as the child was Kell antigen negative.

Conclusion: We report that maternal anti-KELL antibodies were present in breastmilk and were capable of being transferred to feeding child. This persisted at six months of age. If antigen positive, the child could have had an ongoing hemolysis. Breastmilk as a source of antibodies should be considered in children with unexplained anaemia.

2019 May; 13(Suppl 1): S32–S100.

PP 123: Anomalous blood grouping extrinsic to red blood cells mimicking bel phenotype that was not to be!

Background: Discrepancy in ‘forward’ and ‘reverse’ grouping leads to confusion in ABO-typing. Mostly it could be genetic in nature, classified as per the presence and/or absent of antigen on RBCs and antibody in plasma as well as antigen in saliva.

Aim: To study the grouping anomaly in a recently delivered woman who required transfusion.

Methods: A standard protocol, including testing the patient's RBCs with battery of antisera, absorption-elution, and saliva was followed.

Results: The case – A 27 year old female 4 gravida, 3 para required transfusion to correct her hemoglobin. While grouping, her red cells showed group O while serum showed an absence of anti-B. Her RBCs did not react with anti-B or anti-A, B in the battery of anti-sera used. Anti-B was eluted from her sensitized RBCs by anti-B or anti-A, B. Saliva possessed H but no B antigen, indicating to Bel phenotype. The follow up investigation after 2 weeks’ revealed her RBCs group O as was found on previous occasion, but her serum now had anti-B with hemolytic property having an agglutinin titer of 1:512, IgG titer of 1:128 after treatment of serum with 2ME. The problem of missing antibody on earlier occasion was thought to be due to a complement fixing high-titer IgG anti-B that showed an atypical prozone phenomenon due to the deposition of C1qrs macromolecule causing physical distance and thereby preventing the cross linking of the antibody coated RBCs.

Conclusion: The unusual case of erroneous reverse grouping was attributed to complement mediated inhibition of the antibody. One must keep this point in mind while evaluating such rare observation.

2019 May; 13(Suppl 1): S32–S100.

PP 124: Presentation of anti-M reacting at wide thermal amplitude and its clinical/laboratory significance

Background: Anti-M is naturally occurring saline agglutinins that react below 37°C. These antibodies are considered to be clinically significant when they are reactive at 37°C causing HDFN and DHTR. Anti-M appears to be more common in children than in adults and is particularly common in patients with bacterial infections.

Aims: To study the serological spectrum of Anti-M and its clinical/laboratory significance

Methods: This Study was conducted in the Department of Transfusion Medicine, TNMGRMU from Aug 2016-July2018. The group discrepant samples are further evaluated by advanced Immunohematological work-up. If the antibody identified as anti-M, its thermal amplitude was ascertained by tube technique.

Results: In our study, among the discrepant samples, 4 cases of Anti-M identified. Out of 4 patients, 2 were males and 2 were females. One male patient had history of prior blood transfusion and other had no history of sensitization. Both female patients were multigravida and one was presented at 6 weeks of pregnancy. 3 of the four samples were IAT positive. Antibody screening and identification revealed the presence of anti-M antibody. After auto-adsorption, the mono-specific differential card revealed the type of antibody as IgG. Since one of the female patients being antenatal mother, antibody titration was done; anti-M titre is 2. She is on regular follow-up. The other two cases were transfused with antigen negative compatible blood. The remaining one sample without history of sensitization, IAT showed positive reaction at IS and negative reaction in 37oC. However, antibody screening and identification at IS phase revealed anti-M antibody, most probably naturally occurring Anti-M antibody.

Conclusion: Our study has revealed the significance of maintaining M antigen negative blood donor registry for transfusion in emergency situations, even though anti-M is widely accepted as naturally occurring insignificant antibody.

2019 May; 13(Suppl 1): S32–S100.

PP 125: Immunohematological discrepancies - always a single etiology? A case of suspected para-bombay blood group

Background: The parabombay groups have always defied and tested the immunohematologists. These individuals who are H deficient, who might have weak or absent anti-H activity could remain undetected but have serological reactions which can be extremely confounding. We report here a case for whom a parabombay group was suspected.

Case Report: A request was received for a 50-years old female requiring gastroscopy. Her reports from outside showed her blood group to be A positive. Blood grouping done in our laboratory using two different platforms showed forward typing reactions consistent with A positive blood group. However, the discrepancy was noted in the reverse typing, which alongside 4+ reactivity with B cells which was expected, also showed weaker (1+) reactivity with A cells and O cells. However, this positivity was demonstrable in 2 out of 3 A and O cells. The red cells showed a clearly negative reaction with H lectin. The lack of positivity in all A and O cells tested, raised the possibility of a co-existing alloantibody. Antibody screening a performed was negative but reverse typing done at room temperature was positive. A repeat Antibody screen and ID done at 40C showed reactivity consistent with anti-Leb antibody. The patient's phenotype was found to be Lea-Leb-. ABH secretor status was negative. The crossmatch done with 1 Bombay and 5 O red cell units were compatible both at room temperature and coombs phase. The compatible units which were also Leb negative were made available for the patient.

Conclusion: This case highlights the needs to consider multiple etiologies for discrepant serological findings in immunohematology and also the complexities in identifying an alloantibody complicating the patient with a possible Para-Bombay group.

2019 May; 13(Suppl 1): S32–S100.

PP 126: Our experience of ABO-hemolytic disease of newborn - Case series from a tertiary care health centre of North India

Background: ABO-Hemolytic disease of newborn (HDN) is caused by maternal and fetal ABO incompatibility, mainly seen in neonates of blood group A or B born to mothers of blood group O. With the introduction of Rh immune globulin (RhIg) the incidence of Rh D allo-immunization has decreased over last few decades. Consequently hemolytic disease of newborn due to ABO incompatibility and other allo-antibodies have now emerged as major cause of HDN.

Aim: To assess the severity and clinical outcome of ABO HDN in our settings

Methods: Routine investigations are raised from neonatology department after delivery of neonate for direct antiglobulin test (DAT) and ABO blood grouping and are sent to immuno-hematology lab. From May to July 2018, there were 4 cases of ABO–HDN diagnosed. In all cases mother was group O so, serum IgG titre was performed for anti A and anti B. Treatment and clinical outcomes of all the four cases were analysed.

Results: Forward grouping for all cases was B+ve, except one which was A+ve. Mother serum anti A titre were from range 1:512 to 1:1024, similar results were seen for anti B. Titre was also done on eluate of neonate samples which range from 1:128 to 1:1024 for anti A, and from 1:512 to 1:1024 for anti B.

Conclusion: ABO-HDN incidence is 15-17% in Indian population. ABO-HDN should be suspected in neonate with blood type A or B born to mother blood type O with antibody screen negative. Most of the cases of ABO-HDN had benign clinical outcome without the need for exchange transfusion. The intervention of choice was phototherapy, which was also done in the present four cases.

2019 May; 13(Suppl 1): S32–S100.

PP 127: Red cell alloimmunization in multi-transfused patient population: A study from a tertiary care hospital in Odisha

Background: Alloimmunization to red blood cell (RBC) blood group antigens remains a major complication for multi-transfused patients and often presents significant challenges in their medical management. Sensitization to Rh antigens (D, C, c, E, and e) and to K comprise the majority of the RBC antibodies encountered. One major explanation of the high rates of alloimmunization is the disparate distribution of RBC antigens between donors and patients.

Aim: To assess the incidence of RBC alloimmunization in multi-transfused patient population at a tertiary care hospital in Odisha.

Methods: Antibody screening was carried out in 783 multi-transfused patients (thalassemia, Sickle cell disease (SCD) and oncology patients) prior to crossmatching with a commercially available three-cell panel (surgiscreen). Antibody screen-positive samples were analyzed for the specificity of the alloantibody with an eleven-cell identification panel (Resolve Panel A). Above tests were done by Column agglutination technology (CAT) using OrthoBioVue card.

Results: The overall incidence of RBC alloimmunization in multi-transfused patients was 3.19% (25/783), with one case of multiple alloantibodies (anti-c and anti-JKb). Rh alloantibodies were the most common 53.84% (14/26) while Non-Rh alloantibodies being 46.15% (12/26) consisted of MNS 11.53% (3/26), Kidd 7.69% (2/26), Lewis 7.69% (2/26), Lutheran 3.84% (1/26) and unknown antibodies 15.38% (4/26). The highest incidence of alloimmunization was observed in thalassemia patients (2.04%), followed by SCD (0.76%) and oncology patients (0.38%).

Conclusions: Most of the alloantibodies detected in the current study were clinically significant and patients were transfused with antigen-negative crossmatch-compatible blood. Therefore, antibody screening on patients’ samples prior to crossmatching needs to be initiated to ensure safe transfusion practice.

2019 May; 13(Suppl 1): S32–S100.

PP 128: Alloimmunization with multiple allo-antibodies in a pregnant female

Background: Unexpected antibodies in the recipient pose a challenge to blood transfusion services, more so, when multiple allo-antibodies are present. We hereby report a case of a pregnant female with multiple allo-antibodies.

Case Report: A 27 years old pregnant female G2P1L1 with previous caesarean section, presented at 32 weeks of gestation with leaking per vaginum in department of Obstetrics and Gynaecology. Requisition for two units of packed red blood cells was received in blood bank. Patient was typed as AB RhD Negative but discrepancy between cell and serum grouping was observed (cell grouping show AB RhD Negative but serum grouping gave weak positive reaction with A1, B, O pool cells). The direct antiglobulin test and auto-control was negative whereas Indirect Antiglobulin Test was positive. Antibody screen and identification with 3 and 11 cell panel was performed which identified D, C and Jka. Total twelve packed red blood cell bags were typed for D, C, and Jka antigens, out of these only three bags were negative for all three antigens and found compatible with patient's sample. Patient delivered a low birth weight female child by caesarean section but did not require transfusion. Baby was typed as B RhD Positive and direct antiglobulin test was positive. Sample of the baby and father were phenotyped and both were positive for D, C, and Jka antigens. Baby developed jaundice and total serum bilirubin levels reached 20 mg/dl on day 4. Baby was managed with phototherapy and one double volume exchange transfusion with packed red cell unit identified negative for these antigens.

Conclusion: This case highlights the importance of performing antibody screening during antenatal period to detect the presence of clinically significant allo-antibodies.

2019 May; 13(Suppl 1): S32–S100.

PP 130: Anti M: Report of two cases in donor and patient and its clinical significance

Background: Anti M is a common naturally occurring antibody, mostly reactive at temperature below 37 °C and considered clinically insignificant. Sometimes, it is active at 37 °C and may cause hemolytic transfusion reaction and hemolytic disease of newborn. We encountered two cases of anti M in our laboratory. One in a donor presented as Indirect antiglobilin test (IAT) positive and other in a patient presented as crossmatch incompatibility.

Case Report: Case 1 – A 20 yr old male first time voluntary donor was typed as O Rh D negative on cell and serum grouping. Weak D testing was negative, but IAT was 2+ positive. Antibody screen and identification panel (Bio Rad, DiaMed, switzerland) revealed the presence of anti M antibody. Donor was phenotyped as M antigen negative. Anti M detected was reactive at 4 °C with homozygous cells and non reactive at 37 °C and Antihuman globulin (AHG) phase. There was no history of blood transfusion, suggesting it as naturally occurring clinically insignificant antibody. Case 2 – A 3 yr old male child with pyrexia of unknown origin, presented with signs and symptoms of anemia with Hb 2.4 g%. Blood group of the patient was B Rh D positive. Two units were crossmatched and were incompatible. DAT and autocontrol of the patient were negative, but IAT was 3+ positive. Antibody screen and identification panel (Bio Rad, DiaMed, switzerland) showed the presence of anti M. Patient red cells were M antigen negative. Antibody reacted at 37 °C and AHG phase of IAT. Two M antigen negative PRBCs were crossmatched, were compatible and transfused uneventfully. Since there was no history of blood transfusion, it was naturally occurring clinically significant antibody.

Conclusion: Anti M are mostly cold reactive antibodies. Sometimes, may cause HTRs and HDFN. Special immunohaematology card should be issued alerting nature and type of antibody.

2019 May; 13(Suppl 1): S32–S100.

PP 131: Phenotype patient with polytrauma

Background: Correct determination of blood group by forward and reverse grouping is a vital step in pre-transfusion testing. We report transfusion management of a 37-year old polytrauma patient with Bombay (Oh) RhD negative group.

Aims: To determine the actual blood group of the patient and manage his transfusion requirement.

Methods: Blood grouping and antibody titer were done by tube technique. Antibody screen (ABS), direct antiglobulin test (DAT) and compatibility testing were done using gel technique (Bio-Rad, Switzerland). To characterize IgM and IgG di-thiothreitol (DTT) treatment of serum was done.

Results: Forward grouping was O RhD negative, while the reverse grouping showed 4+ agglutination with A, B and O cells. Anti-H lectin reactivity was negative, so the blood group was concluded to be Bombay (Oh) RhD negative. Autocontrol and DAT were negative, and ABS was 4+ (all 3-cells). IgM and IgG titer of anti-H was 1024 and 512, respectively. Before referral, he was transfused multiple O RhD negative PRBCs which might have resulted in hemolytic transfusion reaction. The patient was severely anemic (Hb=4.4 gm/dl) and his renal functions were deranged (urea=220 mg/dL; creatinine=4.9 mg/dL). As the urine output was low (~700mL/day) he was on regular dialysis. Among his first-degree relatives, his brother was found to be OhRhD negative, whose blood was collected and the PRBC transfused to the patient after irradiation. An extensive search was also made through NGO websites and telephonic communications with other blood banks. Two units of OhRhD negative were arranged by an NGO through their registry and air-shipped to our centre. The PRBCs were transfused to the patient without any adverse reaction (Hb=6.8 gm/dl).

Conclusion: Detection of Bombay phenotype requires careful performance of forward and reverse grouping and transfusion management of such rare group patient needs a rare donor registry for ensuring timely availability.

2019 May; 13(Suppl 1): S32–S100.

PP 132: Weaker subtype of “A” detected in a healthy voluntary blood donor

Background: ABO subgroups are phenotypes that differ in the amount of A & B antigen carried on red cells & present in secretions. Several weaker subgroups have been described (eg. A3, AX, Am, A el) which are very infrequently encountered. Here, we present a case of weak A subgroup detected in a 32yr male healthy voluntary blood donor.

Aims: The adsorption -elution and saliva testing plays an important role in resolving ABO blood group discrepancies.

Methods: Both EDTA & clotted samples were collected. Blood grouping done by conventional tube technique. DAT was done by polysp. Gel card method. Adsorption elution done using Human polyclonal anti-A sera. Saliva was tested for secretor status. Anti-A1 titre was performed by pooled A1 cells in double dilution technique.

Results: Blood grouping by CTT method using monoclonal antisera. FORWARD GROUPING showed 1+ reaction with anti-A, negative with anti-B, 4+ with anti-D,4+ with anti-H, 1+ with anti-AB, negative with anti-A1. REVERSE GROUPING showed 2+ reaction with pooled A1 cells, 4+ reaction with pooled B cells, and negative with pooled O cells. Saline control & DAT were negative. The reaction strength with pooled A1 cells persisted up to 4 dilution at room temperature. Adsorption with polyclonal anti-A (three serial adsorption from healthy B group individual) and heat elution (560C for 10 mins) showed agglutination with A cells from six individual donor units on immediate spin. Agglutination-Inhibition testing done on saliva showed ‘H’ secretor only.

Conclusion: Blood group confirmed as weaker subtype of ‘A’. The pattern shows probable blood group is Ax. Since we could not perform the molecular testing, the exact subtype of A could not be confirmed. Adsorption elution and saliva testing appears to be an useful tool to resolve ABO blood group discrepancies.

2019 May; 13(Suppl 1): S32–S100.

PP 133: An algorithm to simplify complex antenatal red cell antibody workup to guide the clinical management

Background: Rh blood group system is most often associated with Hemolytic Disease of Fetus and Newborn (HDFN) predominantly by D antigen. Anti G antibody can cause HDFN either alone or in combination with anti D or anti C. Proper antibody identification and providing a clinical guidance on the fetal outcome can be challenging to the immunohematologist for such cases. This study was aimed to simplify the workup so that timely RhIg can be given to the patient.

Methods: We retrospectively collected data from the blood bank software of all the ante natal antibody screening request over three years. Patients with a positive antibody screening and identification were categorized into pattern suggestive of anti G or others. Anti G gives a pattern of anti D plus anti C in the identification panel. All tests were done using BioRad screening and identification panel in ID system.

Results: Mean age of 30.64 (SD=7.82). The commonest blood group was O negative (39.32%) followed by B negative (34.28%). 66 patient showed reactivity pattern of anti D and 21 showed a pattern of anti D plus anti C or anti G. 3.2% patients were couldn’t rule out the presence of anti E or anti C and the frequency of pattern suggestive of anti G was 11.2%. The complex workup pattern to identify anti G can be simplified by adsorbing onto dCe/dCe cells. The adsorbed sera is then treated with DcE/DcE cells to confirm the presence or absence of anti D (Illustration).

Conclusion: A simple Immunohematological work up can guide the clinician to administer RhIg when the antibody shows pattern of anti G.

2019 May; 13(Suppl 1): S32–S100.

PP 134: Effect of platelet transfusion on thrombocytopenia in patients with dengue hemomorrhagic fever in a tertiary care cenre in Kerala

Background: The proposed study is an analysis of the effect of platelet transfusions on thrombocytopenia in the management of patients with Dengue hemorrhagic fever. Responses to platelet transfusion are quantified using the Corrected Count Increment (CCI) at 1 hour after transfusion and 24 hours. 1st hour CCI above 7500 and 24 hour CCI above 4500 is considered to be evidence of a successful transfusion and 2 transfusions with CCIs below 5000 to 7500 within an hour after transfusion are evidence of refractoriness. Percentage Platelet Recovery calculated both at 1 hour and 24 hours.

Aims: Primary objective – To know the outcome of platelet transfusion in patients with dengue hemorrhagic fever in a tertiary care centre during the period of my study. Secondary Objective – To know the factors responsible forrefractoriness in patients with dengue hemorrhagic fever.

Methods: Study Design – Longitudinal study of effect of increments in platelet counts in patients with dengue hemorrhagic disease who received platelet transfusion therapy. Study Duration: One year six month. Study Subjects: Patients above 14 years, enrolled in the study after taking written informed consent. Sample Size: Out of total 1067 patients suspected of dengue fever since January 2017 upto June 30th 2018, 106 patients confirmed for Dengue, 40 patients had bleeding manifestation, and had underwent platelet transfusion.

Results: The mean age is 41.75 with standard deviation 14.42. Out of total 40 patients, 12.5% patients has refractoriness at 1 hour and 25% for 24 hours. The grade of dengue hemorrhagic fever is statistically significant with refractoriness of CCI 1hour and CCI 24 hour. Out of 22 males 18.2% have refractoriness at CCI 1 hour and 31.8% has refractoriness at CCI 24 hour, not statistically significant. Pretransfusion count and blood group not statistically significant with CCI 1 hour and CCI 24 Hours.

2019 May; 13(Suppl 1): S32–S100.

PP 135: High immunoglobulin-G titer of anti-H in a case of Bombay phenotype

Background: Bombay blood group (h/h or Oh phenotype) is a rare blood group, first discovered at Bombay in 1952. In Bombay Blood group H antigen and Secretor substances are absent due to lack or mutation of FUT-1 and FUT2 genes. Naturally occurring anti H antibody is predominantly IgM in nature and exhibits wide thermal range between 4oC to 37o C and reacts with all red cells except the Oh phenotype. Here we present a case of a 50 year old male with Bombay blood group and high titer of Anti-H IgG.

Aims: To identify the presence of Anti-H IgG in Bombay phenotype and its strength.

Methods: We received a sample in our Immunohematology (IH) lab due to cross-match incompatibility. The blood group of the sample was mentioned as ‘O’ Rh ‘D’ Positive. The IH work-up included Direct Antiglobulin Test (DAT), forward and reverse grouping by Conventional Tube Technique (CTT), antibody screening and antibody titer in saline phase [4°C, room temperature (RT) and 37°C] as well as in anti-human globulin (AHG) phase was done. The serum was also treated with 0.01 (M) dithiothreitol (DTT) to remove the IgM. Then the titer of the DTT treated serum was done.

Results: The saline control and DAT were negative. Forward grouping showed ‘O’ Rh’D’ positive, reverse grouping of the serum showed strong agglutination with pooled ‘O’ cells at RT. There was no reaction with anti-H lectin. Saliva studies showed non-secretor status. The IgM titer of Anti-H was upto 128 serial dilution in RT. Then the serial dilution of DTT treated sera with pooled ‘O’ cell showed 2+ reaction strength in upto 512 dilution in AHG, suggestive of the presence of anti-H IgG of high titer.

Conclusion: Both IgM and IgG immunoglobulins were detected in the serum. The titer of IgG was of clinically significant level.

2019 May; 13(Suppl 1): S32–S100.

PP 136: Severe haemolytic disease of the foetus in an “elevated D’ pregnancy: A rare case report from Western India

The D-/- phenotype is a very rare variant of the Rh system, in which the red cells express D, but no C,c,E,e antigen. These individuals may become sensitized to certain high-frequency antigens of the Rh system through transfusion or pregnancy. Sensitized women carry a major risk of HDFN due to one or more clinically significant antibodies, including anti-Rh27. This presents a major challenge in case management due to unavailability of an antigen negative blood for IUT/ET. A 23 yrs old G2P1L1A0 women with blood group A RhD+ was diagnosed and worked up as a case of HDFN. ICT of the mother was 3+ at 20 wks Period of gestation (POG) with panagglutination in the antibody identification panel (4+) with autocontrol negative. IHL workup revealed the patient's phenotype as RhD+C-c-E-e-(ELEVATED D). Antibody titre was 1:128 with a normal Ultrasound at 20 wks. Serial IHL and radiological workup were carried out on the patient, her relatives (to find compatible blood for IUT), and on the foetus (through cordocentesis) to monitor the severity of HDFN. Cordocentesis done at 24 wks revealed the foetal blood group to be B RhD+C+c-E-e+, DCT 4+, and Hb 2.8 mg%. Ultrasound examination with MCA-PSV at 24 wks indicated a rapid development of fetal anemia and progression to hydrops foetalis. IHL workup of the patient's family revealed blood group A Rh D-/- phenotype in all her siblings (ABOi with foetus). An attempt was then made to find a D-/- compatible blood by working up possible donors from multiple blood banks in the city. However rapidity of progression of foetal anemia and unavailability of a compatible blood for IUT resulted in foetal demise at 27 wks POG. The case suggested that pregnancy in a D-/- woman maybe affected with a rapid progression of hemolysis in the foetus. Timely interventions like dedicated inventories for such rare blood groups at earmarked centers and the possibility of ABOi IUT should be kept in mind to avoid such outcomes.

2019 May; 13(Suppl 1): S32–S100.

PP 137: Drug induced hemolytic anaemia and thrombocytopenia - A case report

Background: Ceftriaxone, a third generation cephalosporin has been reported to occasionally cause drug induced immune hemolytic anaemia (DIHA) and thrombocytopenia (TCP). We hereby report a case where serologic workup of a patient showed ceftriaxone induced DIHA and TCP.

Case Report: A 19 year old male patient operated for gangrenous ileus showed continuous decreasing trend of haemoglobin and platelet count despite multiple transfusions of PRBCs and platelets. All PRBC units issued were compatible in AHG phase (tube method). So, Direct antiglobulin test (DAT) and Indirect antiglobulin test (IAT) were performed using column agglutination test. The patient's IAT was negative. DAT was strongly positive (4+) with polyspecific AHG (anti IgG and anti C3d). After gentle heat elution, DAT was negative. The IAT of eluate from patient's red blood cells (RBC) did not appear to cross react with commercial O pooled RBC's. Detailed history of patient revealed, postoperatively he was on ceftriaxone and piperacillin-tazobactam for last 10 days. So, considering the possibility of DIHA and immune TCP, tests for drug dependent antibodies were performed in the presence and absence of target drugs. Haemagglutination was observed when enzyme treated O pooled cells were incubated with patient's serum, serum sample from healthy donors and 1% suspension of ceftriaxone but no reaction was observed when same mixture was used with 1% suspension of piperacillin-tazobactam. No reactions were detected when pooled serum sample of healthy donors was incubated with RBC's in the presence of both drugs. So in this case antibodies developed in the patient, cross reacted with ceftriaxone. Discontinuation of ceftriaxone resulted in rise in haemoglobin and platelet count of patient.

Conclusion: This case highlights the importance of inclusion of serological tests to determine cross reactivity of drug dependent antibodies in patients receiving any drug as antibiotic treatment inducing immune hemolysis and TCP in the patient.

2019 May; 13(Suppl 1): S32–S100.

PP 138: Past the coombs crossmatch – Blood safety in the alloimmunized

Background: Incompatible transfusion in an allo-immunized individual can result in significant haemolysis. The DGHS requirement of the coombs crossmatch is with the intention of sur mounting this problem. However variability of crossmatch results particularly related to the dosage phenomenon can lead to false-negativity and haemolysis. We report a series of allo-immunized patients and findings on compatibility testing and antigenic phenotyping of compatible units.

Aims: To assess antigenic status of coombs compatible units in patients with clinically significant red cell allo-antibodies excluding anti-D.

Methods: This was a retrospective study including patients with non D red cell allo-antibodies requiring transfusions between March and August 2018. All coombs compatible units were phenotyped for relevant antigen prior to issue. Coombs compatibility and antigen status were compared.

Results: The antibodies identified included 18 non D Rh group, 5 Kidd, 2 Kell, 13 MNS, 6 LeA and 1 duffy. Of a minimum of 5 units cross-matched for each of the 45 patient at least one coombs compatible unit was found. Phenotyping of relevant antigens revealed antigen positivity in 5 of 45 patients (11.2%). Discrepancies were due to antibodies Jka/Jkb/M/s.

Conclusion: While coombs crossmatch provides a definite added layer of safety, it is critical that concomitant phenotyping of red cells for transfusion in the allo-immunized be performed prior to issue. While this could be attributed to the phenomenon of dosage it is interesting to note that no discrepancies were noted in patient's allo-immunized to the Rh system. To prevent a rechallenge and subsequent anamnestic response in sensitized recipients it is imperative that alongside documenting a negative coombs crossmatch that a phenotype is performed to confirm antigen negativity prior to transfusion.

2019 May; 13(Suppl 1): S32–S100.

PP 139: Comparison of two different column agglutination methods for direct antiglobulin test

Background: Direct Antiglobulin Test was conventionally performed using the tube method. Introduction of Column agglutination technology has seen this method being used for Direct Antiglobulin Testing in most labs and is accepted as a reliable method in serological testing.

Aim: The aim of this study is to compare the performance of gel card versus glass bead methods of column agglutination in Direct Antiglobulin Testing (DAT).

Methods: A total of 50 samples for which Direct Agglutination test was requested by the clinician for high suspicion of Autoimmune Hemolytic anemia were included. The tests were done on the blood sample by both the gel card (Diamed; Biorad) and Glass bead method (Ortho Clinical Diagnostics). The test results in both methods were compared based on agglutination grading. Statistical analysis was done on the data collected.

Results: Study is currently in progress.

2019 May; 13(Suppl 1): S32–S100.

PP 140: Analysis of blood components utilisation and quality indicators in cardiac surgical patients in a tertiary care centre

Background: Blood products are one of the most life saving scarce resources among the surgical disciplines. More blood components are required in cardiac surgery compared to most other medical divisions. The overall blood demand depends on the total number of cardiac surgical interventions and complex nature. Due to the awareness of transfusion related immunology by allogeneic blood and its relation to patient outcome, there must be a standardisation of blood transfusion.

Aim: Analysis of blood utilisation statistics and quality indicators in cardiac surgical patients.

Methods: A retrospective analysis of blood components utilisation in Cardiac Surgery unit at the Vinayaka Mission's KirupanandaVariyar Medical College, Salem was done over a period from June 2017 to May 2018. Based on the blood request forms, cross matching and transfusion records, blood components utilisation were analysed. Quality indicators analysed were (1) cross match/transfusion ratio, (2) transfusion probability, (3) transfusion index and (4) maximum surgical blood ordering schedule (MSBOS).

Results: A retrospective single centre study included a total of 224 cardiac surgical cases done by a single unit. Blood components were provided for all of the cardiac surgery patients without fail. A total of 820 units of blood components were requested. The utilisation patterns were PRBC-628 (80.8%), Whole blood-120 (15.4%), FFP-21 (2.7%) and Platelets-8 (1.02%). The population distribution was male 145 (64.73) and female 79 (33.26). The surgical patterns were coronary artery bypass -134 (59.8%), Valve replacement-67 (29.91%) and coronary artery bypass with valve repair-23 (10.26%). The cross match/transfusion ratio-1.05 (<2.0), transfusion probability-87.94 (>30%), transfusion index - -3.46 (>0.5) and MSBOS -5.1.

Conclusion: Usage of blood components by our cardiac surgical division matches the same standard across the globe with minor variations. Regular audit of the blood utilisation practice by our hospital blood transfusion committee helped to match the usage and demand without wastage. In cardiac surgical procedures, adopting various blood conservation strategies by the team of transfusion medicine, surgeon and anaesthetist will help to modify the blood transfusion plans in future.

2019 May; 13(Suppl 1): S32–S100.

PP 141: Retrospective study to find out reasons for discarding whole blood and its components in tertiary care teaching blood bank attach with immuno-hematology and blood transfusion department, government medical college Surat in South Gujarat, India

Background: Blood transfusion is an important constituent of health-care delivery system. Millions of lives are saved every year in regular and urgent situations for medical and surgical indications by the accessibility of safe blood transfusion services. Extension in life expectancy and improvements in medical technology require more and more supply of safe blood for efficient health-care delivery. Human blood has no complete substitute till date. Each unit of blood is precious and has to be utilized properly with minimal discards.

Aims: The aim of this study was to find out the reasons for discarding blood and blood components.

Methods: We retrospectively studied all whole blood and blood components collected during 01/01/2012 to 30/06/2018 at out tertiary care teaching hospital blood bank attach with immunohematology and blood transfusion department, government medical college surat in south gujarat, india.

Results: The total number of blood units collected during this study period was 58,242 & total 6849 (6.05%) components discarded; from this 966 (14.10%) whole blood, 957 (13.97%) red cell concentrate, 2911 (942.50%) fresh frozen plasma, 1507 (22.00%) platelet concentrate, 35 (0.50%) single donor platelets & 26 (0.38%) cryoprecipitate were discarded. the common causes of discarding the blood components were due to expiry date 2283 (33.33%), 1255 (18.32%) were due to seroreactivity for transfusion transmitted infections, leakage of components 1115 (16.28%), low volume 418 (6.10%), hemolysed 32 (0.47%) and other causes were 1746 (25.49%) including components sent for quality control, hyperlipemia, red cell contamination, improper storage, return components, clotted components, etc. Among blood components discarded, most common units were platelets (14.35%).

Conclusion: A properly conducted donor screening, notification and counselling of permanently deferred donors will help in discarding less number of bags which are positive for different tti. Properly implemented blood transfusion policies will help to utilize the blood components in a proper way resulting indiscarding the less number of blood bags due to expiry.

2019 May; 13(Suppl 1): S32–S100.

PP 142: Comparison of quality parameters and biochemical activation markers among buffy coat derived pooled platelets and apheresis platelets (AP-PC) an in vitro study

Background: BCPP have good attributes of both Random Donor Platelets and AP-PC as they are cheaper, contain high count of platelets, can be leukoreduced and be made available in emergency.

Aims: Comparative assessment of physical quality parameters and biochemical activation markers among BCPP (Non-leucoreducedvsLeucoreducedvsLeucoreduced with Platelet Additive Solution [PAS]) and AP-PC.

Methods: Three different preparation of BCPP were prepared – Non-lecoreduced (Part A), Leucoreduced (Part B) and Leucoreduced with PAS (BCPP Part C) using a pool of 15 ABO matched buffycoats to avoid any donor related variations so that each BCPP unit was equivalent to 5 buffycoat units. Ten apheresis platelets were taken as control. Serial samplings were done on day 0,3 and 5 of collection and assessed for volume, platelet count, WBC Count, swirling, pH, Sterility, glucose, lactate, sP-selectin, Intelukin-6, Interlukin-1β and TNF-α.

Results: Comparing the four products BCPP Part A, Part B, Part C and AP-PC, on day 5 the platelet count were 3.42±0.09, 3.34±0.12, 3.31±0.09 and 3.03±0.21 and the difference between BCPP and AP-PC was statistically significant (p<0.05). On day 5, pH was 6.33, 6.42, 6.64 and 6.29 and lactate levels were 172.30, 173.33, 99.02 and 181.91mg/dl respectively showing that biochemical parameters were better maintained in BCPP PAS than with the other products (p<0.001). Inflammatory cytokines were also significantly (p<0.05) higher in non-leucofiltered BCPP part A as compared to leucofiltered products BCPP part B, C and AP-PC; IL-1β was 76.51 in BCPP part A vs 51.60 to 67.89 in others, IL-6 17.86 VS 0.947 to 1.14 in others, TNF-α 27.74 VS 3.78 to 6.05 in others.

Conclusion: This study concludes that leucofiltered as well as PAS suspended Buffy coat pooled platelets are good alternative of conventional platelet preparation and can be very useful in meeting platelet requirements during emergency.

2019 May; 13(Suppl 1): S32–S100.

PP 143: Quality analysis of cellular components obtained by quadruple bag systems using latest automated component extractor

Background: Transfusion of blood components has become an integral part of our health care delivery system. Manual separation of blood components is quite tedious and time consuming. Automated blood component processing by the latest invented technologies not only reduce the workload but also increases the production efficiency with optimized product quality.

Aims: Here we share our experience of analyzing the quality of cellular blood components obtained by quadruple blood bag systems using Automated Component Extractor.

Methods: The prospective study from July 2014 to June 2018 included 556 units of packed red blood cells (PRBC) units and 545 units of random donor platelet (RDP) concentrates. Essential quality parameters of each cellular component described in literature and standards were analyzed and documented in dedicated quality control registers. SPSS software was used for statistical calculations.

Results: The mean platelet volume and platelet yield of RDP concentrates obtained from buffy coat (BC) and evaluated after 3 days of storage were 60.9 ml and 5.33×1010 respectively. The mean pH pf RDP concentrates was found to be 7.1. The mean volume and haematocrit of SAGM suspended PRBCs tested after 21 days of storage were 284 ml and 66% respectively. A one log leukoreduction was observed in >90% of tested PRBCs. All units of RDPs and PRBCs subjected to culture were found sterile.

Conclusion: Automation in blood component preparation significantly enhanced product quality in our blood center. We conclude that quality of blood components is dependent on multiple factors like stringent donor selection, appropriate phlebotomy, strict adherence to standard operating procedures, adoption of automated component separation and proper sampling for quality check.

2019 May; 13(Suppl 1): S32–S100.

PP 144: Optimizing blood component usage in a tertiary care cancer centre: How effective are the initiatives implemented

Background: Requests for blood units are often based on worst case assumptions, resulting in demand for large quantities of blood and wastage of valuable resources. Hence few cost effective interventions were initiated to optimize the use of blood components.

Aim: (1) Calculate the discard rate and reasons for discard of blood components, before and after the interventions. (2) To assess the changes in blood ordering practice by 4 major departments; to calculate CT ratio, Transfusion probability (T %) and Transfusion Index (Ti), before and after the intervention.

Methods: Following interventions were implemented by the department in mid 2017, as part of blood management program. (1) To completely restrict the preparation and usage of Whole Blood (WB) to 100% component preparation and restricting usage of Random Donor platelets (RDP) by preparing High Yield Single Donor Platelets (SDP). (2) To proactively monitor and communicate with 4 major oncology departments to improve their blood ordering practices. Bone Marrow Transplant (BMT), Head & Neck surgery (H&N), Gastro Intestinal surgery (GI), Gynaecologic surgery. Retrospective analysis of data was done one year before & after the intervention by paired t-test.

Results: After the intervention, overall discard rate of Packed red cells/Whole Blood, RDP and SDP were decreased, which was statistically significant, except for SDP. CT ratio increased for BMT whereas it decreased for other departments. The T% and Ti decreased for BMT whereas it improved for other departments, which was significant except for Gynaecologic surgery.

Conclusion: The interventions were very effective in bringing down the discard rates of blood and blood components without any additional cost of operations. An increase in CT ratio for BMT & HL unit was on account of dual policy of type and hold plus type and cross match of specialised blood products.

2019 May; 13(Suppl 1): S32–S100.

PP 145: How do I forestall platelet stockpiling? Experience from a tertiary care center

Background: Among all the blood products, platelets had been reported to have a high rate of outdates due to its unpredictable demand and short shelf life of only 5 days. Researchers have applied techniques of management science and inventory theory to develop a model for inventory management. However, they failed to be implemented due to the variations in the demand and supply and complex computational models.

Aims: To analyze the utilization pattern of platelet concentrates and discuss the method of optimal inventory management.

Methods: We conducted a prospective observational study on platelet inventory practice at our center from January to December 2014. The number of units to be prepared is decided on daily basis by the transfusion medicine faculty or the resident. The utilization, wastage, expiry and the day's cover are calculated for the study period. Future requirement is estimated based on the usage in the previous quarter, discard rare, average increase in usage and an additional 1% for managing disasters.

Results: During this period a total of 6241 and 5706 units of platelet concentrates were prepared and issued respectively. The wastage rate was 5.1% and expiry rate was 3.5%. The average day's cover of platelet units at our centre was found to be 3 days using average monthly stock available and issued platelets. We observed that holding a stock of 45 units of platelets per day we had a cover for about 3 days for issue. Calculation of future requirement (6309) gave a high prediction when compared to the actual platelets prepared (6241).

Conclusions: Understanding and regularly monitoring the inventory, setting up an optimum inventory level, follow of first in first out policy and to have an alternate management plan in times of shortage, like usage of apheresis products are some of the strategies which would benefit in best inventory practices.

2019 May; 13(Suppl 1): S32–S100.

PP 146: Analysis of wastage of blood and components in a teaching hospital in North East India

Background: Among the 10 quality indicators for blood banks by NABH, wastage rate is one of the 5 most important tools. Blood centers, thus necessary to evaluate the wastage/discard rates regularly to improve the inventory management and initiate for appropriate action to improve the services.

Objectives: To evaluate the rates and reasons for wastage of blood and components and to work out strategies to minimize the wastage.

Methods: A retrospective study conducted in the department of Transfusion Medicine, RIMS, Imphal from January 2017 to June 2018. Records of blood units collected from donors, components separated, WB and components issued, discards and reasons thereof etc. during the study period were analyzed.

Results: A total of 17721 WB units were collected during the study period of which 256 (1.44%) were discarded due to TTI reactivity. Forty-two WB units were retained and the remaining 17423 units (99.76%) were separated into components; PRBC=17423 (100%), PC=4920 (28.24%), FFP= 5820 (33.40%) and Cryo=20 (0.11%). The number of WB and components issued were 26638. Total units discarded were 1587 and wastage rate against total units of WB and components issued was 5.95% and against prepared was 5.62%. Component-wise maximum wastage was PC (RDP) with 52.11% followed by PRBC and FFP with 27.73% and 19.10% respectively. Among the reasons, expired date was highest with 1237 (77.95% of all discards), the rest were broken cold chain with 90 (5.67%) and others about 2% each. The discard rate due to expiry of PRBC, PC, FFP and Cryo were 61.59%, 87.4%, 76.2% and 100% respectively.

Conclusion: Our study found expiry units to be most common reason for discard which reveals inadequacies in the inventory management suggesting introspection of the present SOP, need for proper training and educating our staff and rational use of blood by clinicians by generating awareness among the treating doctors regarding appropriate use of each component.

2019 May; 13(Suppl 1): S32–S100.

PP 147: Effect of multiple room temperature exposures for different time periods on hemolysis in stored red cell units

Background: The 30-minute rule that requires discard of red cell units that have been exposed to uncontrolled environments for more than 30 minutes is controversial. Whenever an issued unit is received back it creates a dilemma regarding its subsequent transfusion safety.

Aims: To study effect of multiple room temperature exposures for different time periods on plasma potassium, plasma hemoglobin level and percentage hemolysis in stored red cell units and determine the effect of buffy coat reduction and additive solution on the above parameters.

Methods: A total of 120 bag aliquots of 50 ml each were divided into 4 groups. Groups 1,2 consisted of citrate phosphate dextrose adenine (CPDA) bags while groups 3,4 consisted of buffy-coat reduced citrate phosphate dextrose (CPD) with saline adenine glucose mannitol (SAGM) bags. Exposure to room temperature was given to group 1 for 1/2,2,4 hours on 7,21,28 day respectively and group 3 for 1/2,2,4 hours on 7,21,35 day respectively. Group 2,4 were not exposed. Before and after each exposure samples were tested for hematocrit, total hemoglobin using fully automated cell counter, plasma hemoglobin using Tetramethylbenzidine method and percentage hemolysis was calculated. Plasma potassium was measured using principle of ion-selective electrode.

Results: After consecutive room temperature exposure plasma hemoglobin, percentage hemolysis and plasma potassium were significantly higher in group 1 and 3 on respective days (p<0.001) except for day 7 plasma potassium in group 3. Percentage hemolysis was just below 1% after 2 hour exposure on day 21 in group 1 and 4 hour exposure on day 35 in group 3.

Conclusions: Percentage hemolysis is likely to exceed 1% in CPDA and buff-coat reduced CPD+SAGM bags receiving multiple room temperature exposures when stored beyond 21 and 35 days respectively. The quality of such units should be checked before reissue specially with regards to hemolysis.

2019 May; 13(Suppl 1): S32–S100.

PP 148: Evaluation and validation of two blood collection systems with in-line filters with reference to component quality and leucodepletion process

Background: It is very crucial to evaluate and validate the blood collection and processing system for centres practicing 100% leucodepletion, especially in the light of complex solid organ transplant and bone marrow transplant patients as intended recipients.

Aims: To evaluate the blood bags with in-line filters supplied by two different manufacturers and validate the leucodepletion process and outcome using these two different component separation systems.

Methods: N=118 Compoflow blood bags (Fresenius Kabi) and N=179 Macopharma bags were used for blood component separation as per the manufacturer's instructions. The component volumes, total Hb content of PRBC units, percent recovery of red cells and platelets, platelet yields and component separation time were evaluated. Validation of the leucodepletion process was done by studying the filtration time, percentage loss of red cells from the whole blood collected and residual WBC (rWBC) counts using flow cytometry platform was done.

Results: The volume and Hb content of PRBC units, platelet yield and percentage platelet recovery were higher with Macopharma bags. The time taken for component separation by macopress smart system was lesser (Mean=3 mins 39 secs) than that with component G5 (Mean 4 mins 57 secs) volume of platelet concentrates was higher (Mean 76.4 ml) with Macopharma bags compared to the campoflow bags (Mean 65.5 ml). The compoflow system resulted in lesser (8.5%) red cell loss during in-line filtration than Macopharma system (14.7%). Both bag types yielded rWBC counts <105 in 85% of the units. The time taken for filtration was lesser (<19 mins 26 secs) with Macopharma bags compared to Compoflow bags (24 mins 6 secs).

Conclusions: Both blood bags with in-line filters have advantages and limitations associated with their use. However, both the systems ensure achievement of good component quality and leucodepletion of PRBC units which is required to fulfill special transfusion requirements of transplant patients.

2019 May; 13(Suppl 1): S32–S100.

PP 149: Reasons for wastage of blood and its component in a tertiary care hospital

Background: Blood and blood products wastage is one of key indicator to measure the quality of blood utilization and the efficiency of the transfusion services. According to AABB, quality indicators are specific performance measurements designed to monitor processes and to evaluate adequacy of effective collection processing.

Aims and Objectives: The objective of this study was to determine the rate and cause of wastage of blood and its products in a tertiary care hospital.

Methods: This is retrospective study conducted in Dept of IHBT, DMCH Ludhiana for one-year period from Jan to Dec 2017 to analyse the various factors responsible for discarding of blood and its components.

Results: The total number of blood units collected during the study period was 26476. All components were screened for TTIs. The total number of blood and its component was 64192, out of which 37.6% were packed red cells (PRBC), 31.6% were fresh frozen plasma (FFP). 21.1% were random donor platelets (RDP). 0.4% was cryoprecipitate. The in total discard of blood and its component was 9.8%. The rate of discard was highest for RDP i.e. 29.0% followed by whole blood (11.8%), PRBC (4.8%), FFP (4.6%), cryoprecipitate (4.2%), plasma aphaeresis (3.2%) and platelet aphaeresis (1.4%). The most important reasons for discard of whole blood was TTIs (41.1%) followed by lipemia (29.6%). The majorities of packed red cells were discarded due to TTIs (85.4%) and expired (5.6%). The commonest reasons for discard of FFP were TTIs (70.0%) and leakages (14.1%). The most common reasons for discard of RDP were due to expiry (82.9%) and TTIs (11.6%). The cryoprecipitate (100%) were discarded due to leakage during thawing.

Conclusion: Based on the data collected more than half of wastage is preventable, blood being irreplaceable resourcesneeds to be properly utilizedideally zero percent wastage.

2019 May; 13(Suppl 1): S32–S100.

PP 150: Use of platelet rich plasma for tissue regeneration

Background: Platelet Rich Plasma (PRP) is a platelet product having a higher platelet concentration than in the peripheral blood. PRP & its various forms have become one of the best methods to facilitate the healing process of various tissues due to presence of various types of platelet growth factors. It can be used in disease like chronic non- healing ulcers, chronic joint pain, alopecia, tendinosis/tendinopathy, dental implants, refractive surgeries, cosmetic treatments, etc.

Aim: This study aims at determining effect of PRP & PRP gel on chronic non-healing ulcers & chronic joint pain.

Methods: Informed consent from patients for PRP were received. Clinical features & physical examination findings were noted. Relevant investigations like hemogram, blood sugar levels, pre & post platelet counts were performed. Pain score was recorded via visual analogue scale. PRP was prepared using double centrifugation method & was injected after preparation & PRP gel was applied locally over chronic non healing ulcers & efficacy of therapy was assessed.

Results: Study period-10 months. Total of 62 PRP preparation (38 PRP & 25 PRP gel) for 35 patients- 27 from orthopedics, 3 from Burns & Plastic surgery & 2 from Dermatology. Out of 35- 18 (male) &17 (female). Overall mean age of patients included in this study-39.7 yrs (16-66 yrs). Mean age of males-36.3 (16-66 yrs). Mean age of females-43.4 yrs (20-64 yrs). Mean overall precounts (counts in whole blood sample) of platelet was 2.48 x 103/μltrs. Mean overall post counts (count in PRP preparation) of platelet was 7.36 x 103/μltrs. Out of 35, 1 patient received PRP application 10 times, 1 received 5 times, 1 received 4 times, 4 received twice & 28 received only once. Positive results were obtained after treatment i.e healing of ulcer started/relief in joint pain.

Conclusion: Clinical response after repeat sessions of PRP & PRP gel are satisfactory. Adequate timely follow up proved good results in view of patient satisfaction & treatment efficacy.

2019 May; 13(Suppl 1): S32–S100.

PP 151: Reduction in antibody titers after addition of SAGM - A pilot study on O group prcs

Background: Conventionally, antibodies present in the small amount of retained plasma in PRC units is considered clinically insignificant. Nevertheless, these become relevant in non-identical ABO transfusions especially, preterm low birth weight neonates. In this study we attempted to determine the naturally occurring antibody titres in O group PRC units after the addition of SAGM.

Aims: To determine the antibody titres in O group whole blood & SAGM added PRC units using conventional tube technique. To identify the risk of residual plasma in SAGM added PRC units.

Methods: In this prospective cohort study, O group 350 ml whole blood bags were first weighed and 3 ml samples are collected after proper mixing. Procedure is repeated with PRC prepared from these whole blood bags. Both sets of samples obtained are then centrifuged at 3000 rpm for 3 minutes and the supernatant obtained is subjected to antibody titration at room temperature.

Results: Of the 40 samples analysed, average whole blood volume was 402 ml (SD±31), average PRC volume – 240 ml (SD±20) and average plasma volume – 181 ml (SD±28). Median of anti A and anti B titres in SAGM PRC units was 2 and neat respectively while for whole blood it was 8 and 4 respectively. A significant reduction in mean antibody titres was seen before and after the addition of SAGM (Wilcoxon signed rank test, Z value: -5.334, P value: .05). A small difference in Anti - A/B titres in whole blood & PRC units was also observed, with anti B slightly lower than anti A.(Mann Whitney U test, Z value:-2.351).

Conclusion: Two tube reduction in titre was observed after adding SAGM. However, the use of group OPRC units with low anti-A/B titres is still recommended to increase safety in non-identical ABO transfusions.

2019 May; 13(Suppl 1): S32–S100.

PP 152: A study on influence of donor hematocrit on the procedural parameters of concentrated single donor platelet collected by two apheresis devices

Background: Platelet transfusion remains one of the most important support therapies for thrombocytopenic patients with bone marrow failure. The new generation cell separators and protocols have made it possible to obtain high quality platelets by concentrated SDP (C-SDP) with use of Platelet Additive Solutions (PAS). It is important to assess new procedure performance with regard to cell collection efficiency, collection rate and processing time. The present study was undertaken to know the influence of donor hematocrit on procedural parameters of C-SDP collected by two apheresis devices.

Methods: Retrospective analysis involving 88 and 132 C-SDP procedures by TrimaAccel (65% PAS) and Hemonetics MCS+ (70% PAS) respectively were performed. For studying the influence of donor hematocrit on procedural parameters, donors were categorized into two groups viz Group A (Hct ≤46%) and Group B (Hct>46%). Procedural parameters such as collection efficiency (CE), collection rate (CR), yield per liter (Y/L), yield per hour (Y/H), blood volume processed (%BV) were calculated for both groups and compared between two apheresis devices.

Results: Between the two groups, the mean donor Hct was (43.9% vs 48.6%) and platelet yield (5.6 vs 5.4 x 10e11). In both groups, significant difference was observed between MCS + and Trima equipment in CE (A and B: 53% vs 63%; p<0.05), Y/L (A: 152 vs 183, B: 162 vs 181; p<0.05), Y/H (A: 2.9 vs 5.0, B: 2.8 vs 4.9; p<0.05) and CR (A and B 0.04 vs 0.08, p<0.05). However, we observed %BV processed was significantly higher in donors with lower Hct (74% vs 67%, p=0.01).

Conclusion: Donor Hct does not seem to influence collection parameters except %BV processed between the two equipments. In addition, Trima had better collection parameters when compared to MCS+ for C-SDP procedures.

2019 May; 13(Suppl 1): S32–S100.

PP 153: Study of blood components utilisation in a tertiary care hospital

Background: Transfusion of blood and blood components is an integral part of health care practice. The pattern of blood transfusion has changed considerably in the recent years due to advances in blood banking techniques, prolonged survival and increased frequency of complex surgical procedures. Blood and blood components must be used judiciously and rationally to help minimise the demand supply gap in a blood bank.

Aim: To assess the utilization pattern of blood components (Platelet concentrate) and Fresh Frozen Plasma (FFP) which can help maintain inventory and prevent wastage.

Methods: This study was carried out over a period of six months from January 2018 to June 2018. Necessary data were collected from blood bank registers. Analysis was done for departments from which requisitions were received.

Results: During the study period, total 1058 Platelet concentrate and 536 FFP were issued. Medicine department accounted for the most number of Platelet transfusions, that is 333 (31%) followed by department of Radiation and oncology 187 (17.8%) and Intensive care unit 136 (12.8%). Gastrointestinal Surgery department accounted for the most number of FFP transfusions 101 (19.2%) followed by department of Medicine 94 (18%) and Radiation and oncology 77 (14.6%).

Conclusions: This study highlights the significance of components and usage pattern among different departments. Periodic evaluation of utilization pattern, demand for different blood products also helps to maintain the blood stock.

2019 May; 13(Suppl 1): S32–S100.

PP 154: Single donor platelet-collection efficiency, quality control, and its utilization-an observational study

Background: Single donor plateletpheresis increases the overall yield of platelet collected and decreases the risk of alloimmunisation by decreasing multiple donor exposures particularly in treating haematological malignancies. Providing a better quality apheresis PC will ensure safe transfusion.

Aim: To evaluate the collection efficiency, quality control and utilisation of the single donor Plateletpheresis.

Methods: Retrospective analysis of 62 SDP procedures were done from the period of february 2016 to february 2018 in Tamil Nadu Dr M.G.R Medical University, Chennai, Guindy. Donor selection criteria has been performed as per NACO guidelines. As per demand platelets were collected tested for quality check at the time of issue.

Results: During the study period, 62 procedures were done. Pre and Post procedural donor platelet count were 263×103/μL and 201×103/μL respectively. Average target yield and actual yield achieved were 3×1011/unit and 3.22×1011per unit. Collection efficiency: 51.39%. PC volume were between 200 - 300 ml in 91.93% (n=57), less than 200 ml in 6.5% (n=4) and above 300 ml in 1.6% (n=1) of the procedures respectively. Swirling with score 3 and 2 were present in 90.3% (n=56) and 9.7% (n=6) PC respectively & pH observed between 6.5 & 7.0 at the time of issue. 93.5%of aphaeresis PC met the platelet count 3×1011per unit. WBC contamination is minimal due to leucodepleted filters. All the SDP units collected were utilized. 67.7% (n=42) of SDP concentrates were utilised for hemato-oncological patients 19.35% (n=12) in aplastic anaemia and 12.90% (n=5) due to other causes. Out of hemato oncology cases 85.7% (n=36) were between age of 2-12 and 14.2% (n=6) were aged between 35-50 yrs.

Conclusion: In our study the apheresis PC concentrates were collected with strict adherence to donor selection criteria. The quality parameters revealed good collection efficiency of 51.39%. Due to cost constraints, all SDP concentrates were prepared as per demand and utilised without any wastage.

2019 May; 13(Suppl 1): S32–S100.

PP 155: A retrospective analysis of appropriate utilization of fresh frozen plasma in a tertiary care hospital with reference to evidence based guidelines

Background: Fresh Frozen Plasma (FFP) is mainly used in treatment of coagulation derangements; trauma emergencies. It is the most inappropriately used blood component. Since the guidelines for FFP use in a clinical setting are not well defined.

Aim: To define the appropriateness of use of FFP in the light of its risks and adverse effect. Audit of institute FFP usage with specific aim of assessing appropriate use, based on clinical indication.

Methods: Retrospective analysis of 1846 FFP supplied in 608 patients from the period of January-2018 to june-2018 in tertiary care teaching hospital and the Department of Transfusion Medicine, the Tamil Nadu Dr M.G.R Medical University, Chennai. Detailed analysis of clinical indication and the number of components requested.

Results: 1846 FFP was supplied to 608 Patients. Clinical use of FFP was highest in burns (52%), General Medicine (11.86%), O&G (11.12%), SGE (5.85%), Nephrology (4.22%), Paediatrics (9.15%) and ortho (1.52%). Patients with Deranged Coagulation Profile (DCP) required maximum transfusion of 910 (49.3%) units, Bleeding patients were transfused 644 (34.9%) units and disseminated intravascular coagulation (DIC) 61 (3.3%) units were administered. No information available about diagnosis for 77 (4.2%) units was issued and administered in Emergency Department and 153 (8.3%) units transfused without clear cut indication (hypoproteinemia, volume replacement and patient with normal coagulation profile). Inappropriate requests accounted for 12.5% of the total FFP used.

Conclusions: This study indicates the inappropriate use of FFP to be relatively low in our hospital compared with other studies. FFP is most inappropriately used blood component and should be used judiciously. Regular audit of blood components serves as tool for accomplishment of quality tools and to understand clinical transfusion practices.

2019 May; 13(Suppl 1): S32–S100.

PP 156: Effect of donor variables on the quality of platelet product and comparison of collection efficiency of cell separators

Background: Apheresis in the past was done by manual methods but nowadays cell separators have made it possible to substantially improve quality of apheresis platelets. Platelet yield reflected in transfused platelet dose, influence plt recovery in the patient. This yield is dependent on numerous donor as well as separator related parameters.

Aims: (1) To evaluate the effect of donor variables on the quality of apheresis platelets. (2) To study apheresis platelet preparation by the two cell separators i.e. Hemonetics MCS plus and Fenwal Amicus.

Methods: This was prospective cross sectional study which included 200 procedures; conducted in Dept of IHBT, DMCH, Ludhiana for a period of one year from Jan to Dec 2017. Plateletpheresis procedures were performed on amicus and hemonetics following standard operating procedures. The donors were divided into 5 groups depending on target yield set which was the end point. Platelet yield of the bag and then platelet collection efficiency of the machine was calculated for each procedure.

Results: The effect of donor variables was assessed on platelet yield. Out of 200 donors 198 were male and rest female with mean age of 29.5 ± 8.2 yrs. Donor variables included were age, height, weight, pre donation platelet count, Hb, HCT. Positive correlation (statistically significant) was found between yield and pre donation platelets. Mean platelet yield was 3.67± .81 × 1011. No correlation was seen between age, height, weight, Hb or HCT and yield. Time taken by the procedure was more with amicus but was statistically insignificant. Both cell separators performed procedure with minimum donor discomfort but collection efficiency for amicus was higher than hemonectis.

Conclusion: Donor haematological parameters can significantly affect platelet yield. Therefore, suitable donor selection is vital to the process.

2019 May; 13(Suppl 1): S32–S100.

PP 157: Stability of hemostatic potential of thawed plasma on storage at 2–60C for 5 days

Background: Transfusion of fresh frozen plasma is still an important measure in emergency medicine to prevent disseminated intravascular coagulation after severe blood loss, but thawing procedures can delay its availability. On the other hand, the wastage of plasma, once thawed and not transfused within the defined period, represents an inefficient handling of economic resources. To reduce wastage, we investigated the stability of hemostatic potential of thawed plasma when stored at 2-6 0C.

Aim: To assess the stability of hemostatic potential of thawed plasma when stored at 2-6 0 C for 5 days using APTT, Factors V, VII, and VIII and thrombin generation testing.

Methods: 19 plasma units included in this study were separated from blood collected from the donor and frozen overnight and thawed at 35.80C using plasma thawer. One set of plasma aliquots were stored at -700c and the other set of aliquots from each bag were stored as thawed plasma at 20-6 0 C for 5 days. Factor V, VII, VIII levels, activated partial thromboplastin time and thrombin generation testing were done on first, third and fifth day of storage.

Results: The mean levels of Factor V, VII and VIII of frozen plasma on day 5 of storage were 73.31%, 52.12%, and 62.23% respectively. The mean levels of Factor V, VII and VIII of thawed plasma on day 5 of storage were 67.2%, 50.69%, and 56.97% respectively. The mean change in values of TGT variables were calculated and was comparable between both the groups on day 5 of storage.

2019 May; 13(Suppl 1): S32–S100.

PP 158: A comparative study on the quality of cryoprecipitate prepared by dry and wet method done in a tertiary care centre

Background: Higher utilization of cryoprecipitate in clinical settings propels us to compare methods of cryoprecipitate preparation - Dry (slow thaw) and wet (fast thaw) method.

Aim: To compare the factor VIII and fibrinogen recovery in cryoprecipitate prepared by two different methods of preparation.

Methods: This was a comparative cross-sectional study done in the department of Transfusion Medicine over 20 months. 100 units of whole blood collected at the blood bank comprised the study population. 5 ml plasma was aliquoted& frozen prior to cryoprecipitate preparation for later reference studies. Cryo was prepared using wet & dry methods. The wet method had two different durations for thaw. Factor VIII & Fibrinogen assay was done on the cryoprecipitate and the corresponding aliquoted sample while doing monthly QC. The Recovery (%) of Factor VIII and fibrinogen levels were compared as per gender, season, duration of phlebotomy, blood groups, thawing methods & duration of thaw.

Results: The mean percentage recovery of factor VIII and fibrinogen in wet method were 46.22+13.67 and 50.70+10 respectively showing a better recovery in wet method. The mean percentage recovery of Factor VIII and fibrinogen recovery in dry were 28.66+8.63 and 49.14+ 11.94 respectively. Wet method had two different durations of thawing; 150 & 240 minutes each. The cryo thawed for a period of 150 minutes had better factor VIII and fibrinogen recovery (%) when compared to the 240 minutes thaw.

Conclusion: The recovery of factor VIII and fibrinogen in cryo was better when wet method of thawing was used. The 150 minute thawing duration for the wet method was better compared to the 240 minutes of thawing. A standardized protocol in processing of cryo has to be improvised considering other variables like blood group of the donor, duration of phlebotomy, the duration of thawing and seasonal variations.

2019 May; 13(Suppl 1): S32–S100.

PP 159: Prevalence of transfusion transmitted infections in voluntary and replacement donors

Introduction: Transfusion of diseases is one of the major hazards of blood transfusion. In India we screen blood units for TTI namely HIV, HBV, HCV, Syphilis & Malaria. Accurate estimate of risk of TTIs in donor samples gives an idea of epidemiology of these diseases in community.

Aim: To determine the prevalence of TTI in voluntary and replacement donations.

Materials and Methods: This retrospective study was conducted in SUM Hospital blood bank Bhubaneswar, Odisha from Jan 2013 to June 2018. A total of 46,125 blood units from blood donors were tested for TTI by ELISA (Qualisa-Qualpro Diagnostics- TULIP). Test were performed according to manufacturer's instructions. All the reactive samples were tested in duplicate before labeling them seropositive. The donated unit was discarded when found positive for any TTI.

Results: A total of 46,125 donors were included in the study. Of these 38,264 (82.9%) were voluntary and 7,852 (17.10%) were replacement. Male donors 45,074 (97.7%) outnumbered females 1051 (2.3%). A total 686 (1.4%) of the 46,125 donors tested reactive for TTI out of which replacement donors were 558 (1.2%) and voluntary 128 (0.2%). The overall seropositivity for HBV was 434 (0.9%) out of which replacement were 308 (0.66%) and voluntary 126 (0.34%). HIV had seropositivity of 207 (0.4%) with replacement 142 (0.3%) and voluntary 65 (0.1%). HCV constituted 41 (0.08%) of which replacement donors were 34 (0.07%) and voluntary 7 (0.01%). Seropositivity of syphilis was 4 (0.08%) with equal positivity in both group of donors and none tested reactive for malaria.

Conclusions: Study shows that prevalence of TTI was high in male replacement donors. Seroreactivity was higher for HBV followed by HIV, HCV, syphilis and malaria. Stringent donor screening, encouragement for voluntary donation and retention of voluntary donors should therefore be prioritized.

2019 May; 13(Suppl 1): S32–S100.

PP 160: Enhancing blood safety through nucleic acid amplification testing: Experince from a tertiary hospital blood bank from Eastern India

Background: Transfusion-transmitted infections are a major problem associated with blood transfusion. Nucleic acid amplification testing (NAT) is not yet obligatory in India for blood donor screening. The primary benefit of NAT is the ability to reduce residual risk of infections due to its short window period.

Aims: Here we share our 5 years experience of screening our blood donors by The Roche cobasTaq Screen MPX platform.

Materials and Methods: All the non reactive blood donations tested by CLIA between 23 November 2013 and 30 July 2018 were included in the study. NAT for HBV-DNA, HCV-RNA and HIV-RNA in the minipool of 6 samples was performed using the Roche cobasTaq Man MPX assay. Sample positive for hepatitis- B virus (HBV) DNA was further screened for anti-HBc antibody & antibody to HBsAg (anti-HBs).

Results: Of the total 48342 blood donations during the study period, anti-HCV, HBsAg and anti HIV by CLIA were detected in 264 (0.55%), 234 (0.48%) and 121 (0.25%) donors respectively. A total of 47723 samples were tested for NAT of which 19 (0.04%) donors were found to be carriers of HBV DNA each with a viral load of <6IU/ml. HIV RNA was detected in one donor who was otherwise non reactive for anti HIV. The NAT yield was observed to be 1 in 2386 donations. Eleven (58%) donors carrying HBV DNA were sero-reactive for anti-HBc and 9 (47.4%) showed reactive anti-HBs antibody.

Conclusion: Introduction of NAT has successfully identified the pre-seroconversion infectious blood donors and occult hepatitis B. Despite its cost effectiveness issues NAT can prove to be a standard of blood donor screening in the future.

2019 May; 13(Suppl 1): S32–S100.

PP 161: Seroprevalence of hepatitis E virus among blood donors from a tertiary care center in North India

Aim and Background: US Food and Drug Administration (FDA) has recognized the Hepatitis E virus (HEV) as a transfusion-transmissible infectious agent in the year 2004. The information on the prevalence or transfusion risk from India is limited. Therefore, we conducted a study to find the seroprevalence of HEV infection in blood donors and transfusion risk by detection of IgM HEV antibodies by ELISA and HEV RNA by Reverse Transcription Polymerase Chain Reaction (RT- PCR).

Materials and Methods: The study was conducted at our tertiary care centre from September 2016 to December 2017. One thousand and three donors who donated during the study period were tested for the IgM antibodies against HEV using ELISA. Of these, 256 random donors were subjected to HEV RNA detection by the RT-PCR method which included HEV IgM reactive donors by ELISA and ELISA non-reactive donors selected randomly.

Results: In our study 8 (0.8%) donors were reactive for IgM antibody against HEV by ELISA. Mean age of these donors was 36 years (27-54 years). Of them, 3 were B positive (0.8%), 3 were O positive (0.9%) and one was A positive (0.4%). All donors reactive for HEV ELISA were negative for HIV, HBsAg, HCV by CMIA and non-reactive by ID-NAT. Two donors who were reactive for HEV ELISA were positive for HBcAb by CMIA. None of the 256 donors tested was reactive by RT-PCR.

Conclusion: There was 0.8% seroprevalence of HEV IgM antibody in our population. Further large-scale studies to detect the HEV prevalence in Indian population should be conducted.

2019 May; 13(Suppl 1): S32–S100.

PP 162: Seroprevalence of transfusion transmissible infections among blood donors in regional institute of medical sciences hospital, Imphal

Background: Blood transfusion is considered a life-saving procedure. But it also carries a potential risk factor for transmission of serious infections like HIV, HCV, HBV, Syphilis etc. Thus, strict mandatory screening of TTIs as per national guidelines is necessary for safe clinical blood transfusion.

Aim: To find out the seroprevalence rate of TTIs in blood donors.

Methods: A retrospective study was carried out in the Department of Transfusion Medicine, RIMS, in which data about the blood donors donated during the period between January 2017 and June 2018 were collected. The data of the donors including the seroprevalence were collected from various relevant registers and were analysed. The TTI tests included in the study were antibody to HIV (I and II), anti-HCV, and HBsAg and RPR (for syphilis) tests. The tests had been performed by using 3rd generation, NACO approved ELISA and RPR test kits.

Results: A total of 17,721 blood units were collected during the study period, of which 5326 (30%) were voluntary blood donors (VBD) and 12395 (69.9%) were replacement donors (RBD). The overall TTI reactivity was 256 (1.44%), and the order of seroprevalence were HCV (0.66%), HBV (0.53%), HIV (0.20%) and syphilis (0.03%). The TTI reactivity among VBDs and RBDs were 54 (21.09%) and 202 (78.9%) respectively. Among VBDs, HCV (0.45%) had highest seroprevalence rate followed by HBV (0.33%), HIV (0.18%), Syphilis (0.037%) and the rates in the RBDs for HIV, HBV, HCV, Syphilis were 0.21%, 0.61%, 0.75% and 0.04% respectively. Co-infections were found in 8 (0.04%) donors, all were RBDs.

Conclusion: The present TTI reactivity rate, which was more in RBDs; and presence of co-infections, needs introspection for improvement especially in the donor screening, and emphasis on increasing VBDs would ensure blood safety.

2019 May; 13(Suppl 1): S32–S100.

PP 163: Socioeconomic impact of the predonation hepatitis B surface antibody testing in the blood donors

Background: India has an estimated prevalence of 3% HBV carrier rate that is more than 37 million HBV carriers at current population level. It is not uncommon for these HBsAg carriers to present as blood donors. However, studies in the past indicate that since they are clinically asymptomatic they may not return for a counseling session later on. An important opportunity for their contact testing and further management of these high risk individuals is thus lost.

Aims and Objectives: To study the economic and social impact of pre-blood donation HBsAg testing on the HBV carrier counseling.

Materials and Methods: All the blood donors who cleared the preliminary questionnaire and found fit for further testing before blood donation were tested using whole blood rapid card test kit for HBsAg. Comparison was made with the past HBsAg reactive blood donors (reactive for HBsAg after blood donation) who were called for counseling before the study was started.

Results: There was no difference in the overall prevalence of HBsAg reactivity in the study (4440 donors) Vs the control (1205 donors) population (2.34% Vs 2.32%; p=0.74). However, with the pre-donation HBsAg testing, a significantly higher proportion of the reactive donors could be counseled as against the standard post donation counseling (104 Vs 28 donors; 92.3% Vs 39.3%; p<0.001). Moreover, in addition to the significantly reduced exposure of the blood center staff to the HBsAg reactive blood, the blood center incurred 63% less expenditure in terms of the cost of blood collection.

Conclusion: Pre-donation HBsAg testing of the blood donors significantly increases the counseling opportunity of the asymptomatic HBV carriers in the community. Such a testing decreases the potential exposure of the healthcare staff to HBV reactive samples and also results into a significant cost saving for the blood center.

2019 May; 13(Suppl 1): S32–S100.

PP 164: An analysis of discordent result of chemiluminscence serology and ID nucleic acid amplification testing for infectious disease markers

Background: To reduce risk of transfusion transmitted infection Serology testing is supplemented by NAT. Discordant results in NAT & serology makes the testing strategy challenging. Therefore it is important to further evaluate the discordant samples to understand the safe testing strategy.

Aim: To evaluate the discordant samples NAT & Chemiluminscence in order to suggest suitable & safe TTI testing protocol.

Materials and Methods: We collect nearly 40000 units annually from voluntary donors. Serology testing is done by Vitros® 3600 by Ortho Clinical Diagnostics & NAT by Procleix® Panther System by Grifols. Further analysis of all discordant samples (Sero yield & NAT yield) was done. All the discordant samples were repeated by another Chemiluminescence system (Abott) Architect & Viral load PCR was also done. In addition, Western Blot for HIV & anti HBc for HBV was also performed.

Results: From January 2016 to March 2017, total 50443 samples tested for Chemiluminscence& NAT. Out of 50443 samples, 357 samples (0.7%) were reactive by Chemiluminescence and 150 samples were reactive by both Chemiluminscence & NAT. Sero Yield was 207 & NAT Yield was 14. Out of 62 discordant samples of HIV, only one was reactive with another chemiluminescence test, all were negative for Western Blot & PCR Viral load. Out of 16 discordant samples in HBV, all were negative with second chemiluminescence, 5 were positive for anti HBc (Occult infection) and none has PCR Viral load. Out of 141 discordant samples in HCV, 21 were reactive with another chemiluminesce and viral load was not detected with any sample.

Conclusion: Addition of NAT in Blood Screening helps to detect window period donations and occult Hepatitis B infections and our NAT yield found is 1 in 5000. We found good correlation in Serology & NAT in HBV but high Sero-yield in HCV.

2019 May; 13(Suppl 1): S32–S100.

PP 165: Significance of implementing individual donor nucleic acid testing to minimize the residual risk of transfusion transmitted human immunodeficiency virus, hepatitis C virus and hepatitis B virus infections – A study among blood donors in a government tertiary care teaching hospital in Chennai

Background: The primary aim of Blood Transfusion Services is to provide blood and blood components to the recipients as safe as possible. Even with the development of highly sensitive and specific serologic assays, Transfusion Transmitted Infections (TTI) still pose a threat to blood safety.

Aim: The objective of this study was to assess the significance of implementing individual donor NAT (ID-NAT) for HIV-1&2, HCV and HBV to minimize the window period donations.

Materials and Methods: A total of 7320 donations from JAN to AUGUST 2018 were tested for all three viruses using enzyme-linked immuno sorbent assay (HIV Ag-Ab, HCV-Ab and HBsAg by ERBA LISA) and ID-NAT using ProcleixUltrio Elite assay (Grifols). All initial NAT reactive samples and serology non-reactive were tested in triplicate and NAT discriminatory assay for HIV-1 and 2, HCV and HBV were performed.

Results: Out of the 7320 samples, 14 (0.19%) were found to be ID-NAT reactive but seronegative. All these 14 samples were reactive for HBV by discriminatory assays and nonreactive for other two viruses. The NAT yield rate was 1 in 523. Quantitative viral load assay of these NAT yield samples revealed a minimal viral load ranging from 6 IU/ml to 70 IU/ml suggesting very early period of infection.

Conclusion: 42 TTIs (14 x 3 components) have been prevented by implementing IDNAT testing in our Blood Bank within the above period. After observing the above results, the prevalence rate is highly alarming and safety of blood transfusion can only be improved by strict donor selection, quality assured sensitive serological methods and introduction of universal NAT testing. ID-NAT testing for HIV-1 & 2, HCV and HBV can significantly improve the efficacy of screening and it is an additional layer of blood safety by preventing preseroconversion window period donations thus reducing the treatment cost and burden on healthcare.

2019 May; 13(Suppl 1): S32–S100.

PP 166: Changing trends in incidence of transfusion transmitted infections

Background and Aims: Blood bags collected in India are mandatorily screened for HIV antibodies, HBV surface antigen, HCV antibodies, Syphilis (RPR) antibodies and Malarial parasites which are major reason for discard of blood units. The incidence of TTI in donor population undergoes changes as the incidence of infections changes in general population because of preventive measures and disease control. The aim of the present study is to study the changing trends in incidence of TTI in donor population which indicates the current trend in general population.

Materials and Methods: TTI screening results obtained at the blood bank at Government Villupuram Medical College and Hospital from the year 2010 to 2017 were collected and analyzed to watch the changing trends in incidence of TTI.

Results: A total of 26,572 blood units were collected during the period from 2010-2017. The overall screening results also showed a declining trend from 97 in 2012 to 52 in 2017 in spite of increasing trend in blood collection. HIV declined from 4 reactive in 2012 to 1 reactive in 2017. HBV declined from 91 reactive in 2012 to 50 reactive in 2017. HCV declined from 10 reactive in 2010 to 1 reactive in 2017. Syphilis declined from 1 reactive in 2010 to nil in 2017. Malaria declined from 1 reactive in 2011 to nil in 2017. Thus HIV, HBV, HCV showed a declining trend over the years.

Conclusion: Strict adherence to donor questionnaire and better donor screening has led to deferral of doubtful donors with risk of disease transmission. Deferral of high risk donors, follow-up of seropositive donors with referral to appropriate specialty department and strict deferral of febrile donors help to reduce the incidence of TTI and thus better blood inventory.

2019 May; 13(Suppl 1): S32–S100.

PP 167: Seroprevalance of transfusion transmitted infections in multiple transfused thalassemia major patients

Background: Thalassemia also known as “Cooley's anaemia” is an inherited disease of the red blood cells classified as a haemoglobinopathy. It is characterised by decrease or absent synthesis of normal globin chain. Appropriate and regular red cell transfusion remains the main treatment of choice for a large number of patients with thalassemia major. These patients who are maintained on transfusion regimen can develop various complications due to multiple transfusions, one of them being transfusion associated infections.

Aims: To study the rate of seropositivity to Human Immunodeficiency Virus (HIV), hepatitis B and C infections among patients with β-thalassemia major receiving multiple transfusions.

Methods: The study was performed from January 2016 to June 2018 in DMCH, Ludhiana on 189 multi-transfused thalassemia patients. All the patients were screened for HIV (p24 antigen & antibodies to HIV-1 & HIV-2), Hepatitis B (Hepatitis B surface antigen) and Hepatitis C (Anti-HCV antibodies) by ECLIA (Electro Chemiluminescence Immunoassay) COBAS e 411 ROCHE. Further screening and discriminatory assays by PCR (Polymerase Chain Reaction) confirmed the presence of TTI's.

Results: Out of the 189 multiple transfused patients, 54 (28.6%) were infected with TTI's. HCV was positive in 52 cases (27.5%), HBV in 1 case (0.5%), and HIV in 1 case (0.5%). PCR confirmed the presence of HBV DNA in 1 case (0.5%), HIV RNA in 1 case (0.5%) and HCV RNA in 22 cases (11.6%).

Conclusion: HCV was the leading TTI in multitransfusedthalassaemia major patients in this study. Provision of safe and adequate blood supply to these patients is a key to improve their quality of life and longevity. Transfusion associated diseases are overcome by safe donor selection and, further, by application of better screening methods.

2019 May; 13(Suppl 1): S32–S100.

PP 168: Study of nucleic acid amplification testing yield in elisa tested nonreactive blood units at indu voluntary blood bank, Vadodara

Background: Risk of Transfusion Transmitted Infection (TTI) is always there during any transfusion. Nucleic Acid Testing is one of the best currently available technology when used in combination with ELISA to reduce the window period and risk of TTI.

Aim: To study the NAT yield & determine the benefit of NAT technology as an additional safety for Blood Transfusion services.

Methods: We used fourth generation ELISA to screen the blood samples of all voluntary and replacement blood donors. All ELISA negative samples were tested by MP-NAT based on the principle of POLYMERASE CHAIN REACTION using COBAS 201s with MPX v2.0. All NAT reactive donors were retested for viral load by COBAS TAQMAN 48.

Results: Starting from 19/05/2018 to 31/08/2018, MP-NAT was performed on 7143 donor samples which were found non reactive on ELISA testing. Out of the 7143 donors tested by MP-NAT, 6 were found to be NAT-reactive which were ELISA non reactive (NAT yield) for HBV. The prevalence of NAT yield cases among was 1 in 1190 donations tested (0.084 %). Out of 6 NAT reactive donor samples, 5 were tested for quantification test. All those 5 blood units were having viral load above the detection level & 3 out of 5 donors samples were having viral load more than 6IU/ML & 2 were having <6IU/ML Since we supply blood components (packed red cells, fresh frozen plasma and platelet concentrate), these 6 units of blood would have yielded 18 components & hence 18 patients could have been infected with HBV viruses.

Conclusion: In the vast majority of blood units tested, the results of ELISA and MP-NAT for HIV-1&2, HBV and HCV were concordant. MP-NAT did detect the presence of viruses missed by ELISA in some blood units. Its widespread use in blood banks would ensure safer blood transfusion.

2019 May; 13(Suppl 1): S32–S100.

PP 169: Prevalence of nucleic acid amplification positivity in seronegative blood donors: A retrospective study in a tertiary care hospital

Background: Transfusion of blood and blood components, as a specialized modality of patient management saves millions of lives worldwide each year and reduces morbidity. It is well known that blood transfusion is associated with a large number of complications, transfusion transmitted infections (TTI) is one of them. Nucleic acid Amplification (NAT) test has been implemented in many developed countries as a screening test of blood donors to reduce the window periods of viruses.

Aim: To detect the seroprevalence of Hepatitis B (HBV), Hepatitis C (HCV) and Human Immunodeficiency Virus (HIV) in all seronegative blood donors and providing the information regarding blood safety.

Methods: A retrospective review of donor records within periods from July 2016 to June 2018 was done in the Department of Transfusion Medicine, SCB Medical College & Hospital, Cuttack, Odisha. All the samples were tested for Hepatitis B Virus (HBV), Hepatitis C Virus (HCV) and Human Immunodeficiency Virus (HIV1/2) first by Enzyme Linked Immunosorbent Assay (ERBA elisa, 3gen) and all seronegative ELISA samples were retested by Minipool-NAT.

Results: A total of 51,627 donors were screened for Hepatitis B (HBV), Hepatitis C (HCV) and Human Immunodeficiency Virus (HIV1/2) by ELISA. Out of which 51,109 seronegative ELISA donors were retested by MP- NAT. The overall ELISA positivity and NAT positivity were 1.01% and 0.09% respectively. The prevalence NAT positivity was 0.086% in voluntary and 0.093% in replacement donors. The NAT positivity of HBV, HCV and HIV1/2 were 0.08%, 0.003% and 0.006% respectively. NAT yield was 1 in 1100 seronegative donors.

Conclusion: Transfusion transmitted infections (TTI) screening is a primary concern of blood safety. This study reveals replacement donations are more unsafe in comparision to voluntary. Based on the results we feel that to reduce the risk of these infections, non-remunerated voluntary donor services need to be instituted along with adoption of more sensitive methods of TTI screening for improvement of blood safety.

2019 May; 13(Suppl 1): S32–S100.

PP 170: Comparison of nucleic acid amplification testing and routine serological testing in patients undergoing hemodialysis

Background: Patients on chronic hemodialysis are at high risk for contracting bloodborne infections like Hepatitis C Virus (HCV), Hepatitis B Virus (HBV) and Human Immunodeficiency Virus (HIV). Blood transfusions can also increase this risk. Routine serological tests like Enzyme Linked Immunoflourescent Assay (ELFA) are done at regular intervals to detect the occurrence of HCV/HBV/HIV infection in these patients. Use of Nucleic Acid Amplification Testing (NAAT) will enable us to identity these infections much earlier and reduces the transmission of infections among patients in dialysis units.

Aims: To study the ability of NAAT testing to detect HIV, HCV and HBV infection compared to the currently used serological tests in hemodialysis patients.

Methods: Patients undergoing hemodialysis for minimum period of six months at our centre were tested randomly for HBV DNA, HCV RNA, HIV RNA by ID-NAAT test. Serology tests were done every three months and this data was collected from their medical records. The risk factors for infection like dialysis at multiple centres, blood transfusions, repeat use of dialyser were recorded.

Results: Eighty eight patients (62.5% Male and 37.5% female) were analysed. Mean age of 60 ± 13.49 years. 12.5% of patients had undergone dialysis at more than one centre. Average number of blood components transfused per patient was 10.8 units. Duration of dialysis ranged from 13-151 months. Dialyser for each patient was used multiple times after sterilisation. All patients were negative by serology (ELFA) and ID-NAAT.

Conclusion: ELFA assay seems to be equally effective as NAAT in detecting HCV, HBV and HIV infections in hemodialysis patients. Proper Sterilisation protocols, designated machines for TTI positive patients, dialysis at single centre and transfusion of NAAT screened blood could have prevented TTI in our hemodialysis patients.

2019 May; 13(Suppl 1): S32–S100.

PP 171: Utility of multiple platforms to increase the sensitivity of transfusion transmitted diseases (malaria and syphilis) screening in blood donors – A study from hinduja hospital and MRC Mumbai

Background and Aim: Transfusion Transmitted Disease is a major threat to blood recipients. With various sensitive methods and techniques, the risk of Malaria & Syphilis transmission has drastically reduced. Factors associated with this are effective donor selection, donor counseling, method and sensitivity of screening tests. With this background we carried out a prospective study to compare sensitivity of the different methods used for Malaria and Syphilis testing and results compared with gold standard.

Methods: Prospective study was carried out between January 2017 -August 2018. In Malaria testing 70 (42 positive & 28 random negative) donor's samples and for Syphilis 73 (45 positive & 28 random negative) samples were tested. For malaria Pan Malaria card, Malaria Pan/Syphilis Combo (Immunochromatography), peripheral smear and for Syphilis Redgen kit (flocculation test), Malaria Pan/Syphilis Combo and Treponemapallidumhemagglutination test (TPHA) tests were carried out. All the positive samples were tested on all the platforms.

Results: 28 (40%) out of 70 donors were negative for malaria in all three platforms. 42 (60%) donors had positive results in either of the platforms. 34 (80.96%) of 42 had all three platform positivity and 8 (19.04%) tests had discordant results. Analysis showed 7/8 cases were positive on Pan Malaria & peripheral smear and only 1 (2.39%) test was positive on both immunochromatography platforms however negative on peripheral smear examination. 28/73 (38.35%) donors were negative for Syphilis in all platforms. 45 (61.65%) donors had positive results in either of the platforms. 31 (68.89%) of 45 had all three platform positivity and 14 (31.11%) tests had discordant results. Analysis showed 9/14 cases were positive in both rapid tests however negative on TPHA. Remaining 5 cases were positive in Pan/Syphilis combo kit & TPHA and negative on Flocculation test (Redgen).

Conclusion: This study highlights the importance of identifying sensitive screening tests to reduce risk of TTI.

2019 May; 13(Suppl 1): S32–S100.

PP 172: Quality control of HIV, hepatitis B surface antigen and hepatitis C virus screening kits - Discordant results observed between ELISA and rapid card test

Background: Quality control (QC) is simply to ensure that the results generated by the tests are correct and to monitor the accuracy & precision. ELISA (Enzyme linked immunosorbent assay) is a recommended and preferred screening technique for blood banks. Many blood banks use rapid, easily performable and user friendly kits.

Aim: To evaluate the discordant results observed between ELISA and Rapid card test for quality control of HIV, HBsAg & HCV screening kits.

Materials and Methods: A prospective study carried out in the Department of Transfusion Medicine, the Tamil Nadu Dr M.G.R Medical University in the month of July -2018. QC of ELISA for HIV, HBsAg & HCV is done by using in-house control and virotrol with Levey-Jennings Chart (LJ chart). QC of Rapid card test for HIV, HBsAg and HCV by in-house control and virotrol are also done to observe the concordant and discordant results of quality control test between ELISA and Rapid card.

Results: In Elisa, the westgard rule is applied with± 2SD, in total 20 runs of QC kits were analysed for three infections, in house control ± 2SD,20 runs cut off value for HIV, HBsAg & HCV are 0.11-0.82, 0.2-0.46 & 0.35-0.67 whereas in QCrun values are 0.21-0.8,0. 21-0.4 & 0.39-0.63 respectively. In virotrol± 2SD, 20 runs cut off value for HIV, HBsAg & HCV are 0.48-0.88, Nonreactive & 1.47-2.9 whereas in QC runvalues are 0.58-0.73, Nonreactive & 1.8-2.7 respectively. Hence no discordant results found in all the three infections by using In house control whereas virotrol shows discordant result for HBsAg. In Rapid card test, the in-house controls show Nonreactive for all the three infections whereas virotrol shows Nonreactive for HBsAg.

Conclusion: Failure of the rapid kits to detect HIV, HBV & HCV reactive samples may be due to Inadequate coating of the antigens, Nature of the antigens used and Genetic heterogeneity of the virus. It signifies the need of ELISA testing in TTI to ensure the safety of blood and blood products being issued to the needy patients.

2019 May; 13(Suppl 1): S32–S100.

PP 173: Prevalence of viral markers among blood donors at a tertiary care centre - A retrospective study

Background: Blood transfusion is associated with several risks particularly exposure to blood transfusion-transmissible infections (TTI), including: Hepatitis B virus (HBV), Hepatitis C virus (HCV), Human immunodeficiency virus (HIV), syphilis, malaria and others. The safety of donated blood can be estimated by monitoring the prevalence of viral markers in the donor population.

Aim and Objectives: The aim of this study was to evaluate the prevalence of viral markers among blood donors.

Materials and Methods: Over a period of three and a half years (January 2015 to July 2018, a total of 40,253 blood units were collected from healthy voluntary and replacement blood donors. The donated units were serologically screened for hepatitis B surface antigen (HBsAg), antibody to hepatitis C virus (anti-HCV) and HIV.

Results: A retrospective analysis of blood donors’ records covering the study period was undertaken. The records were analyzed to evaluate the prevalence of TTIs. The prevalence of HBsAg was highest in the year of 2016 and 2017 (1.6%) compared to 2015 (1.5%) and 2018 (1.1%). There was a marked decline in the prevalence of HCV infection from 0.03% in 2015 and 2016 to 0.01% in 2017 and no case detected in 2018 till July. The prevalence of HIV was highest in the year 2016 (0.17%), 2017 (0.15%) compared to 2015 (0.06%) and 2018 (0.07%).

Conclusions: The study reveals that the decrease in HBV, HCV and HIV prevalence among blood bank donors might be associated with the introduction of immunization programs and an increased health awareness throughout the country. However, there is a need for continuous surveillance of TTIs.

2019 May; 13(Suppl 1): S32–S100.

PP 174: Prevalence of transfusion transmitted infections among voluntary blood donors with different ABO, Rh and Bombay blood group system

Background: Transfusion of unscreened blood is associated with risk of Transfusion transmissible infection. Genetically determined ABO blood group antigen may prevent binding of possible causative organism to polysaccharide on cell and non secretors of antigens are at risk of TTI.

Aim: To determine the prevalence of TTI among voluntary donors with ABO, Rh and Bombay blood group system.

Materials and Methods: This was a retrospective study conducted at blood bank of The Tamilnadu Dr.MGR Medical University and Tertiary care hospital over a period of five years from January 2013 to December 2017. Details were collected from the records in the Transfusion Medicine department.

Results: Out of the 41,013 donors, overall 532 (1.3%) were showed seropositivity. Among ABO system, “A” blood group (1.5%, 116/7864) showed comparatively more prevalence of TTI than other groups. Highest percentage of HBsAg was in blood group “A” (1.28%, 101/7864) followed by “B” (1.23%, 173/13971), “AB” (1.1%, 31/2858), and “O” (0.1%, 163/16319). Percentage of HCV was more in group O (0.89%, 14/16319) followed by group AB (0.14%, 4/2858). VDRL was commonly observed in group AB (0.04%, 1/2858) followed by group B (0.03%, 4/13971). Highest percentage of MP antigen was in group B (0.007%, 1/13971) followed by group O (0.006%, 1/16319). Donors with AB group (0.07%, 2/2858) showed slightly higher percentage of HIV infection. The prevalence of TTI seropositivity was relatively higher among Rh-negative blood donors (1.56%, 35/2232) comparing to Rh positive donors (1.28%, 497/38781). HBV (1.3%, 29/2232) & HCV (0.27%, 6/2232) seropositivity percentage was highest in Rh Negative. Bombay blood group showed no positivity for TTI.

Conclusion: In our study, we observed only marginal variations in TTI prevalence among ABO, Rh and Bombay Blood group systems. Hence, further studies to be conducted with larger number of donors for conclusive remarks.

2019 May; 13(Suppl 1): S32–S100.

PP 175: Markers for transfusion transmitted infections in a medium sized blood bank

Introduction: One of the adverse effects of allogenic blood transfusion is the transmission of infectious agents. Florence Nightingale, more than 100 years ago said “No stronger condemnation of any hospital or ward could be pronounced than the single fact that zymotic (infectious) disease has originated in it, or that such a disease has attacked other patients than those brought in with them”. It should, therefore, be obligatory on those who are involved in transfusion of blood to apatient for saving his life, that the blood transfusion does, no harm to the patient. Nothing could be worse than the fact that in an attempt to save life, blood & blood products having transmissible infectious agents have been given. 25-30% of multiple transfusion recipients in india show evidence of infection with both HBV and non A and non B hepatitis or HCV. To be licensed, blood banks must screen each donor unit for HBsAg, antibodies to HIV-1 and HIV-2, HCV and VDRL.

Objectives: (1) To study and calculate the percentage of HIV, HbsAg, HCV and VDRL positive donors out of total voluntary and replacement donors in 2017 (2) To reduce risk of post transfusion infections in future.

Methods: In 2017, 17798 voluntary and replacement donors at this centre were screened for the above markers, using commercially available kits.

Results: The annual data was tabulated month wise. There were 11.13% HbsAg positive donor units, HCV antibodies were detected in 0.22 % of total donors, annual VDRL reactivity was 0.37% and HIV was detected in 2.14% of donors tested.

Conclusion: Prevention is the key to handling transfusion transmitted infections. The Implementation of donor selection, sensitive screening tests and effective inactivation procedures can ensure the elimination of risk of acquiring transfusion transmitted infection with the collaboration of national haemovigilance system for protecting a secure blood product supply

2019 May; 13(Suppl 1): S32–S100.

PP 176: Blood donor notification and counseling: our experience from a tertiary care hospital in South India, Puducherry

Introduction: An adequate, safe supply of blood and blood components is a crucial part of blood transfusion services in order to avoid transfusion related infections. Blood donors with reactive screening test results are informed of their results by telephone call, and are requested to come for counselling either at the blood centre or the integrated counselling and testing centre. Many notified donors either do not respond at all or do not follow up their first visit to the blood bank. This study was undertaken to determine the response of blood donors after notification of their reactive status by telephone call.

Aim and Objectives: To evaluate the response rate of transfusion-transmissible infection (TTI)-reactive donors after notification of their abnormal test results during first half of 2018.

Materials and Methods: This is an observational descriptive study performed in our department over a period of 6 months. We evaluated the response rate of TTI-reactive donors after notification of their abnormal screening test results using telephonic intimation.

Results: During the study period 85 blood donors were found to be seroreactive. Of these 85 seroreactive cases, 1 was HIV positive, 80 were reactive for Hepatitis B surface antigen (HBsAg), 3 were Hepatitis C (HCV) positive and 1 was VDRL reactive. Among the TTI-reactive donors (85), 43 (50.5%) were contacted telephonically. Of the 43 contacted donors, the response rate was 32% as only 14 donors reported for one to one counseling.

Conclusion: Donor notification and post-donation counseling are an essential aspect of the blood bank that entails provision of information on serological status, assess the impact of test results on the donor and finally referral for medical care.

2019 May; 13(Suppl 1): S32–S100.

PP 177: Study of incidence of hepatitis C virus in leukemia patients attending cancer institute, Chennai, India

Aim: A retrospective study was carried out to determine the incidence of Hepatitis C Virus [HCV] Infection among Leukemia patients undergoing treatment at Cancer Institute, Chennai.

Introduction: When transfusion is an essential part of treatment protocol, the screening of donor blood and itscomponents for transfusion transmitted infections like HCV assumes great significance.

Materials and Methods: The retrospective study was carried out at Cancer Institute, Chennai, India during the period 2009 – 2015 on leukemia patients undergoing treatment. The incidence with respect to number of transfusions, demographic profile, time gap between detection of leukaemia and HCV seroconversion, genotype, viral load, Liver function tests and co infection was analysed.

Results: A total of 79 cases were detected HCV seropositive (Genotype 1). Mean age of presentation was 13.60 ± 11.09 years, 64.1% male, mostly belonging to low income group from rural areas. The average number of total transfusions received by these patients before seroconversion was 7.35 ± 3.77 units and mean time gap between diagnoses of leukemia to HCV seroconversion was found to be 13.42 ± 18.50 months. 43% of these patients reported abnormal liver function parameters. The incidence proportion of HCV seropositive cases decreased significantly from 62.5 cases per 1000 population (2009) to 7.17 cases per 1000 population (2015).

Conclusion: In this study, Seropositive HCV infection was found primarily in young male leukemia (ALL) patients from low socio economic and rural background. The incidence proportion of HCV seropositive cases showed a decrease when transfused with donor blood screened for HCV by Ag-AB Monolisa Ultra V2 as compared to screening with Anti-HCV Assay.

2019 May; 13(Suppl 1): S32–S100.

PP 178: Prevalence of hypothyroidism, diabetes mellitus and delayed puberty in patients of thalassemia major in a tertiary care center of Jammu Province, Jammu Kashmir, India

Background: Thalassemia is a common genetic disorder which is associated with a lot of complications. Frequent blood transfusions result in increased iron deposition in various tissues leading to dysfunction of many organs of our body. Endocrine disorders constitute a major part of such complications increasing the morbidity of thalassemia manifold in the affected patients.

Methods: This is a prospective study carried out in 64 thalassemia major patients attending thalassemia day care centre at SMGS Hospital Jammu from December 2014 to November 2015. Patients were examined and investigated for presence of one or more endocrine disorders including diabetes mellitus, hypothyroidism and delayed puberty.

Results: Endocrine disorders were detected in a total of 22 patients. Diabetes mellitus was detected in 4.7% (n=3) patients, hypothyroidism in 4.7% (n=3) patients and delayed puberty was found in 26.6% (n=17) patients. Mean serum ferritin level was found to be 2885.5 ng/ml and there was no significant difference in patients affected with endocrine disorder and those without any endocrine disorder.

Conclusions: Endocrine complications occur commonly in patients of thalassemia major. Increasing life span of thalassemia patients has increased the number of patients living with these disorders. A lot of morbidity occurs due to the presence of one or more of these disorders. Hence timely detection of these disorders by screening in all patients of thalassemia should be done to initiate treatment at the earliest so as to limit the morbidity caused by these disorders.

2019 May; 13(Suppl 1): S32–S100.

PP 179: To evaluate blood transportation workflow efficiency by implementing irreversible temperature monitoring device in a blood bank of a tertiary care centre

Background: Wastage of Packed Red Blood Cells (PRBCs) due to interruption in cold chain during transportation hampers the blood transfusion services in meeting its goal of safe and efficient supply of blood.

Aim: We undertook this prospective and observational study with the aim of finding out whether there is any difference between dedicated trained transportation team and patient's relatives in cold chain maintenance of PRBCs.

Methods: Pre validated irreversible temperature monitoring strips (Time strip®) were attached to PRBCs before issue from the blood bank. We included 120 PRBC units into two groups of 60 each; one with trained carriers (dedicated staff) and the other with untrained carriers (patient's relatives). Time of issue from blood bank, time of receipt at the recipient's end, total time elapsed prior to starting transfusion and Time strip® reading were recorded and compared.

Results: Statistically significant results were obtained between the two arms (Trained Vs Untrained) with p value <0.002 on Chi square testing). 73% of PRBCs were delivered within 30min in the trained group, when compared with 45% in the untrained arm. No statistically significant results were obtained in the average time elapsed from the time of issue to the start of transfusion in both the groups. 6 units were discarded in the untrained arm due to unacceptable Time strip® reading.

Conclusion: This study reiterates the importance of dedicated transportation team in maintaining temperature of PRBCs to less than 10°C during transportation and time limit of 30 min between the issue of blood & starting transfusion, thereby minimizing chances of hemolysis which may cause transfusion reactions due to improper handling of blood, especially in a tropical country like ours.

2019 May; 13(Suppl 1): S32–S100.

PP 180: Evaluation of blood requisition forms and utilization practices at a tertiary care hospital transfusion medicine department in Kerala

Background: Quality of a blood service is evaluated in three analytical phases (pre analytical, analytical, and post analytical). Pre analytical phase accounts for 68% of total errors. The pre analytical phase includes hospital-based procedures outside the domain of blood bank such as requisition form filling, proper sample identification etc. The Blood Request forms (BRF) is the first line of communication between the clinician and transfusion medicine. Auditing of blood transfusion requests and calculation of quality metrics (cross match-to-transfusion ratio [C:T] ratio, %T, TI) have been considered the most effective way of evaluating the appropriateness of transfusion.

Aims: (1) To audit the Blood Bank request forms so to evaluate its appropriateness. (2) To assess the utilization of the blood product by Calculating the number of quality metrics: that is the Cross match transfusion ratio (C:T ratio) transfusion probability (%T), and Transfusion index.

Methods: A cross-sectional, prospective study was conducted. The blood request forms received from Jan 20 to Feb 2018 were evaluated for its completeness. The red cell concentrate utilization practices were assessed using C:T ratio, transfusion probability (%T), and TI using equation.

Results: Among 350 requisition forms, 221 (63%) were for males and 129 (37%) females. Only (2.5%) requisitions completed all parameters, majority were incomplete. Percentage of parameters which remained incomplete were, patients name 4%, Age: 10%, Sex: 7%, Hospital ID: 2%, Ward: 3%, Bed Number: 7%, Patient group and Rh: 5%, Diagnosis: 5%, Indication: 6%, previous transfusion: 11%, referring doctors name: 4%. Referring doctors signature: 47% and If female history of pregnancy or still birth: 17%. The Cross match Transfusion ratio was assesses 1.15 and transfusion probability, 89.3%and transfusion Index, 1.5 which shows appropriateness.

Conclusion: Incomplete requisition forms may at times cause alarming issues, need of continuous education programme. Small proportion of blood product marked as urgent was issued in urgency, thereby, compromising inventory management. There was appropriateness in the utilization as the quality metrics were under normal limits.

2019 May; 13(Suppl 1): S32–S100.

PP 181: Systematic root cause analysis of blood bag damage in component laboratory

Background: An efficient error reporting system can limit the magnitude and severity of incidents and prevent future episodes. The MERS-TM approach was applied to an incident involving damage of blood bag during centrifugation which occurred in component laboratory using systematic root cause analysis (RCA) and subsequent implementation of corrective and preventive action (CAPA) was done.

Methods: Systematic RCA was performed followed by a fish bone diagram. After identifying the cause, incident was analysed with the aid of MERS-TM and all four evils of error were assessed. Adequate corrective action was undertaken and strict adherence to preventive measures was ensured.

Results: The incident occurred in component laboratory where the technician noted damage of blood bag while taking it out from the centrifuge buckets after the initial hard spin. A tear was found in the bottom of blood bag which seemed to have been caused by some sharp object/s, while the ports were found intact. The damage was extensive with leakage being sufficient enough to fill the buckets with blood. It was brought to the notice of resident doctor posted in the laboratory who initiated an RCA followed by CAPA for the same. Suspicion of possibility of balances which were moderately sharp in nature and or sealed end of tubing which were accidently kept on the bottom of blood bags were raised. Subsequently technicians were reiterated about the importance of adherence to standard procedures to avoid occurrence of any such incidents in future. Further, resident and technician on duty were assigned the responsibility of supervising correct placement of blood bags into the buckets.

Conclusion: Correct placement of tubing of blood bags in the buckets prior to centrifugation can help circumvent wastage of precious blood resource. Active reporting of such incidents and analysis using systematic RCA approach must be encouraged.

2019 May; 13(Suppl 1): S32–S100.

PP 182: Are physicians utilizing blood appropriately? A retrospective study of transfusion practices in a tertiary care hospital

Background: Excessive cross matching in addition to being wasteful of resources has adverse consequences on the management of blood inventory and blood quality. Cross match/to transfusion (C:/T) ratio is an important measure that is used to assess how the physicians are utilizing the blood transfusion services. A C:T ratio of >2 indicates excessive ordering of cross matched blood.

Aims: The main aim of this study was to analyse the pattern of blood cross matching and transfusion requests requirements by the physicians with the aim of creating updated local policies that minimize resource wastage.

Methods: 76 months of retrospective data from may 2012 to august 2018 was collected which included RBC cross match requirements requests and all RBC units transfused.

Results: A total of 40,908 units of packed red blood cells were cross matched and 24,787 units were used. The overall C:T ratio was 1.65 corresponding to 60.59% of red cell usage and 39.41% of wastage. The Obstetrics and Gynaecology department had the highest C:T ratio of 2.8 followed by surgery department with 2.2. The department of medicine had the lowest C:T ratio of 1.2 followed by department of oncology with 1.4.

Conclusions: The primary outcome of the study was compliance of overall C:T ratio with the international guidelines. We found that current deficiency of MSBOS (Maximum Surgical Blood Order Schedules in the departments like obstetrics and gynaecology and surgery departments were the major factor responsible for high C:T ratio. Therefore MSBOS were suggested for common elective procedures in both the departments.

2019 May; 13(Suppl 1): S32–S100.

PP 183: Audit of blood requisition and utilization in elective neurosurgical procedures

Background: Over ordering of blood components is common in major cardiac and neuro surgeries. These leads to unnecessary burden on transfusion services in form of wastage of blood components, manpower and financial outflow. So continuous audit of blood transfusion during major surgeries ensures better usage of resources, cost effectiveness and streamline the process.

Aims: The study aims to analyse the differences in number of packed red blood cells (PRBC) requested, cross-matched and transfused in patients’ undergone neurosurgery in tertiary care institute.

Methods: This retrospective observational study includes 450 patients undergone elective neurosurgery for 6 months period from January 2018 to June 2018. Data were collated from the blood bank E-record. The data collected include patients’ age, sex, type of surgical procedure, number of PRBC units requested, cross matched, returned, transfused, cross match to transfusion ratio (C: T), transfusion index (Ti) and transfusion probability (%T).

Results: During study period for 450 patients, 1628 units of PRBCs were requested of which 1118 (68.6%) units were cross-matched, while 954 (59%) units issued, and only 330 (20.2%) units were transfused. The overall C: T ratio, transfusion probability (%T) and transfusion index (Ti) were 3.38, 39.11% and 0.73 respectively. Maximum PRBC transfusion were in craniotomy and excision surgical procedure {66.36% (219/330)} while minimum transfusion was in cranioplasty procedures {0.12% (4/330)}. The overall difference between PRBC cross matched to issued and PRBC issued to transfused were found statistically significant (p=0.000).

Conclusion: This study highlights wide gap between ordering of blood and transfusion. This indicates the need of implementing Maximum Surgical Blood Ordering Schedule (MSBOS) for elective neurosurgical procedures to reduce wastage of blood components, save time and manpower of transfusion services and efficient management of blood inventory.

2019 May; 13(Suppl 1): S32–S100.

PP 184: Impact of school awareness program on increasing voluntary nonremunerated blood donors in the population - Study from a tertiary care centre in South India

Background: Access to safe blood is a key component of effective health care and voluntary non remunerated blood donation has been universally declared to be the cornerstone of safe blood.

Aims: The present study was undertaken to determine existing awareness regarding blood donation among students and local stakeholders and to design specific methods to increase awareness among these groups.

Methods: A close ended questionnaire was administered to 22 teachers from 4 different schools and their responses were compiled and analyzed. This was followed by an awareness training programme following which the same questionnaire was administered after the intervention to assess if there was an improvement in knowledge and attitude towards blood donation. 154 school students and 36 parents were also included the study and the same study algorithm was followed. Focused group discussion based on knowledge of blood and blood donation, myths of donation and ideas to improve blood donation in the community was also assessed.

Results: There was a significant improvement in knowledge regarding blood donation among all participants post the training session. However with respect to attitude scores, there was an improvement in attitude towards blood donation among teachers and students. More than half of the teachers (64%) had not donated previously and there was no correlation between demographic variables and knowledge of blood donation. Children were aware of the need of blood, but was unaware of the concept of voluntary blood donation. Fear of needles, deleterious effects on health and lack of awareness were the most common reasons for non-donation among all groups.

Conclusion: This study highlights that greater knowledge about blood donation does not transform into actual practice. It is essential that donor education and sensitization through extensive campaigning strategies need to be initiated to target different population groups and to reinforce motivational perceptions.

2019 May; 13(Suppl 1): S32–S100.

PP 185: The pattern of red cell utilization in four major specialties in a government medical college and hospital of Eastern India

Background: The tussle between the daily demands of blood and the supply realized is never ending. There is always a limitation to our stores of safe blood. So, it is imperative that the supply and utilization graphs remain at least somewhat proportional.

Aim: To assess the proportion of red cell units that are not being put to optimal use or are being wasted.

Methods: Four departments, with major blood requirements, were randomly selected, viz., Gynaecology and Obstetrics (G&O), Paediatrics, Medicine and Surgery. They were monitored for 16 days. On the 17th day, a surprise audit was made at the in-patient department and assessed for the number of units transfused and the number of units still present in their inventory.

Results: In the department of G&O, 156 red cell units were issued whereas only 47 units were transfused and 34 units were still in their refrigerator. In Paediatrics department, 289 units were issued versus 206 transfused and in the Medicine department the figures were 274 and 187 respectively. Out of 177 units issued to the Surgery department, 50 were transfused and 66 were in their domestic refrigerator. In G&O, the cross-match to transfusion (C/T) ratio was 3.32, transfusion probability (%T) was 27.11% and Transfusion Index (TI) was 0.39. In Paediatrics department, C/T ratio, %T and TI were 1.4, 71.75% and 0.77 respectively. In Medicine department, C/T ratio was 1.47, %T was 69.62% and TI was 0.79. In the department of Surgery, C/T ratio was 3.54, %T was 34.59% and TI was 0.37.[ C/T ratio <= 2.5, TI >= 0.5 and %T >= 30%, implies significant blood usage].

Conclusion: There appears significant underutilization/wastage of red cells mainly in the G&O and Surgery departments. This is detrimental to the overall blood stores, thus depriving many in need of this essence of life.

2019 May; 13(Suppl 1): S32–S100.

PP 186: Type and screen protocol versus coombs crossmatching in hospital transfusion practice

Background: Pre transfusion compatibility testing mainly includes ABO & Rh typing, screening the patient's serum for unexpected antibodies and cross matching. If antibody screen is negative and the patient has no previous history of sensitization, then 99.9% of ABO compatible red cell units would be compatible in Coombs cross match.

Aims: (1) To compare the safety of Type and Screen protocol and Coombs cross matching for compatibility testing. (2) To estimate the efficiency of blood utilization in the department of Obstetrics and Gynaecology for elective surgical procedures.

Methods: This was an observational study implemented in three phases on 1800 requisitions from the department of Obstetrics and Gynaecology for crossmatching from December 2015 to December 2017. In Phase I, on 150 requisitions Coombs crossmatching was performed as the pretransfusion test. In Phase II, on 150 requisitions, Type and screen protocol was performed. In Phase III, on 1500 requisitions Coombs crossmatch and Type and Screen protocol were done independently without considering the result of each other. The safety, cost and turnaround time of both the protocols were compared. Blood utilization statistics were estimated.

Results: In the present study, T&S protocol gave safety level of 100% when compared with Coombs crossmatch. It was detected that 30% of the technologist's time could be saved by using T&S protocol. The usefulness of T&S protocol was shown through detection of unexpected antibodies in 0.4% of cases, which would have been missed otherwise. The transfusion rate amongst the blood ordered and crossmatched were 5.37% and 10.09% respectively. On implementing the T&S protocol; the Crossmatch to Transfusion ratio, Transfusion probability, Transfusion index were corrected from 14.9, 11.3%, 0.13 to 1.1, 100% and 1.07 respectively.

Conclusion: A review of blood ordering habits and blood utilization statistics along with implementation of T&S protocol can help in improving the hospital transfusion practices.

2019 May; 13(Suppl 1): S32–S100.

PP 187: Evaluation of errors in blood requisition forms

Background: Errors can take place in any section of a blood bank. An essential section is the reception of requisition forms. The errors present in the received requisition forms can be fatal for the patients if not noticed on time. Therefore, a system is required to monitor the errors or non-conformance in the blood bank continuously. In this context, we evaluated the request forms received at our blood bank. In the evaluation, we found different possible errors in approximately 10% of the forms.

Methods: Being a stand-alone blood bank, we receive around 12400 requisition forms from different hospitals in Siliguri and nearby area every year. We evaluated all the requisition forms and samples (37288) collected between the year 2015 and 2018 for detecting errors and non-conformance.

Results: We were able to find 3946 erroneous requisition forms out of which 3207errors were minor errors for example, incorrect/missing name (80/12), missing sex or age of the patient (220), unstamped form (1230), unsigned form (410), unmentioned clinical diagnosis (840), incorrect or missing ward/bed number (383) and unmentioned hospital name (44). While 739 errors were severe errors for example, samples found with wrong group (49), unmentioned blood group (250), different registration number on form and sample (364), unmentioned required component name (58) and sample received was of another patient (18). These errors could have been fatal to the patient but our efforts and preparedness detected all the erroneous requests on time.

Conclusion: Approximately 10% requisitions were erroneous among which 20% (2% of total) were severe. The evaluation reports were shown to the hospitals and they were convinced to modify their SOP in order to minimize the errors by rechecking the forms and samples at the hospital level before sending to the blood bank.

2019 May; 13(Suppl 1): S32–S100.

PP 188: Estimating blood needs for a tertiary care hospital

Background: There is always a disparity between demand and supply of blood units in hospital. Even though there cannot be a single standard or formula for estimating blood needs and estimation needs to be tailored to the local region.

Aim: The aim of the present study was to estimate the blood needs of the population attending a tertiary care hospital.

Methods: Blood needs were estimated by two methods and compared at the blood bank of Government Villupuram Medical College and Hospital, taking the blood utilization pattern of 2017. In the first method, discarded units during the same period, 10% for expected increase in demand and 4% for disaster management were added to 2017 blood utilization data. In the second method, 6 months data of blood usage was obtained (July to December 2017); one week of high usage was subtracted and the sum total was divided by 25. This gave the blood needs for each week.

Results: In the first method, number of units utilized from January to December 2017 was 8799 blood components. 378 blood components were discarded of which 77 units were red cell units. After computing for discard and demand, the estimated blood needs is 10375 blood components. In the second method, average weekly demand was calculated to be 181.61. When computed for a year (52 weeks), the expected demand was 9445 blood components. Assuming 60% component separation, 5247 blood units have to be collected.

Conclusion: Whatever the formula used, the local blood needs have to be estimated. This will help to assess the burden for blood bank, recruit adequate blood donors and advocate appropriate use of blood and blood components.

2019 May; 13(Suppl 1): S32–S100.

PP 189: A study on blood requisition and transfusion practise in a tertiary care hospital

Background: Blood loss is an integral part of any surgical procedure. It depends upon type of surgery, coagulation status & surgical expertise. Demanding large quantities of blood of which little is used commits exhaustion of valuable time and resources.

Aims: (1) To determine Cross match Transfusion ratio, Transfusion probability and Transfusion index. (2) To determine how appropriately blood is ordered in various surgical departments.

Methods: A total of 412 cases, which had request for PRBC cross match for various surgical procedures during May 2018 were studied retrospectively with respect to number of units ordered, cross matched and transfused. Then CT ratio, Transfusion probability and Transfusion index were calculated.

Results: CT ratio was 4.6 for elective cases & 2.9 for emergency cases. It was high for Surgery, Orthopaedics, Obstetrics & Gynaecology departments and acceptable level for Cardiothoracic surgery & Neurosurgery department. 96.5% of the requests from Obstetrics and Gynaecology department was appropriate whereas 3.5% was inappropriate.

Conclusion: CT ratio of less than 2.5 is associated with significant blood usage. In our study it was much more than 2.5 except Neurosurgery and Cardiothoracic surgery departments. This indicates unnecessary cross matches. In surgeries which have insignificant blood loss, cross match can be avoided.

2019 May; 13(Suppl 1): S32–S100.

PP 190: Evaluation of transfusion - related adverse events in thalassemic patients: A study from tertiary care centre at Baroda

Introduction: Regular blood transfusion is the mainstay treatment and a life saver for thalassemia patients, it may be associated with various transfusion related complications.

Aim: This study was carried out to evaluate immune and non- immune transfusion reaction in thalassemia patients over a period of one year at SSG Hospital, vadodara.

Methods: This study was carried in the Department of ImmunoHematology and Blood Transfusion, Govt Medical College Baroda from May 2017 to April 2018. A total of 100 thalassemia major patients were included in study who received regular blood transfusions. A detailed history, examination and monitoring of patients throughout the transfusion and up to one week after transfusion for any reaction was done. Various investigations like serum ferritin, blood sugar, liver function test, thyroid function test, serum calcium level, ELISA for HBsAg, HCV, HIV, malaria and syphilis were done. Antibody screening and identification was carried out in all patients.

Results: There were 27 females and 73 males in total of 100 patients. Adverse transfusion reaction rate was 5.47% (93/1700). Allergic reactions constituted 13%, febrile reactions 25%, FNHTR 51% and febrile with allergic constituted 11%. Antibody screening revealed 3 patients positive for alloantibody and 5 patients positive for autoantibody. TTIs were found sero-reactive in 5 patients. Serum ferritin levels with multi-organ dysfunction were found despite chelation in 3 patients. There were 2 patients of diabetes mellitus. Total 25 patients who were more than 16 years of age, 11 (44%) patients had delayed pubertal development and hypogonadism.

Conclusions: In the present study, it was found that thalassemia patients suffer from numbers of complications owing to repeated blood transfusion. Thus meticulous monitoring of transfusion and highly accurate, sensitive and specific investigation must be carried in these patients to minimise chances of adverse transfusion complications.

2019 May; 13(Suppl 1): S32–S100.

PP 191: Do certain hla expressions increase the risk of psoriasis an Indian expereince

Background: Transfusion Medicine is a multispecialty that includes immunology. This study focuses on HLA association in Psoriasis since studies showing HLA association in Psoriasis are mostly from the West with very little information about HLA association in Psoriasis in the Indian Population. Psoriasis is an immune mediated genetically determined skin disorder affecting 0.5 – 3.0% of the Indian population. Psoriasis is a multi-factorial disease and has been associated with certain HLA expressions.

Aim: To screen Psoriasis patients for HLA - A & B and determine its association.

Methodology: The study was conducted in the Department of Transfusion Medicine at a tertiary care hospital in South India. Hundred Psoriasis patients (cases) and 100 healthy blood donors (controls) were enrolled in the study. Samples from the Psoriasis patients were collected from Dermatology Out Patient Department and the samples from the controls were taken from voluntary blood donors in the Department of Tranfusion Medicine. HLA typing was done using PCR – SSP method. The HLA results were analysed stastically by open epidemiology and SPSS software and its association with Psoriasis was determined.

Results: The alleles which were found in higher frequency among the cases were HLA A*02 (45% of the cases), HLA A*11 (34% of the cases) and HLA A*24 (35% of the cases). HLA B*35 was found in 36% of the cases.

Conclusion: Certain HLA alleles are present in higher frequency in the disease population than the controls implying that individuals expressing these alleles may have a higher relative risk of developing Psoriasis. The findings of this study on HLA association with psoriasis differs from previous studies done on different ethnicities.

2019 May; 13(Suppl 1): S32–S100.

PP 192: HLA-C association in psoriasis

Background: Transfusion Medicine is a multispecialty that includes immunology. This study focuses on HLA association in Psoriasis since studies showing HLA association in Psoriasis are mostly from the West with very little information about HLA association in Psoriasis in the Indian Population. Psoriasis is an immune mediated genetically determined skin disorder affecting 0.5 – 3.0% of the Indian population. Psoriasis is a multi-factorial disease and has been associated with certain HLA expressions.

Aim: To screen Psoriasis patients for HLA- C and determine its association.

Methodology: The study was conducted in the Department of Transfusion Medicine at a tertiary care hospital in South India. Hundred Psoriasis patients (cases) and 100 healthy blood donors (controls) were enrolled in the study. Samples from the Psoriasis patients were collected from Dermatology Out Patient Department and the samples from the controls were taken from voluntary blood donors in the Department of Tranfusion Medicine. HLA typing was done using PCR – SSP method. The HLA results were analysed stastically by open epidemiology and SPSS software and its association with Psoriasis was determined.

Results: The alleles which were found in higher frequency among the cases were HLA C*06 (52% of the cases) and HLA C*07 (33% of the cases). HLA C*12 was found in 35% of the controls.

Conclusion: Certain HLA alleles are present in higher frequency in the disease population than the controls implying that individuals expressing these alleles may have a higher relative risk of developing Psoriasis. This study confirms the previous studies findings that HLA – C*06 is strongly associated with psoriasis.

2019 May; 13(Suppl 1): S32–S100.

PP 193: A pilot study of prevalence of Rh, MNS, kell, duffy, kidd and P1 blood group antigen in blood donors in Western Rajasthan

Introduction: Transfusion support is one of the most widely used therapy in hospital practices but have many risks which can be fatal. Still, RBCs for blood transfusion are mostly only matched for the major antigens, ABO and D, an approach except in chronic transfusion recipients (e.g. thalassemia), who additionally match for minor antigens.

Aim: Objective of this pilot study is to know blood group antigenic frequencies in Western Rajasthan population.

Materials and Methods: The study was conducted at Blood Bank, attached to department of Transfusion Medicine, Sardar Patel Medical College &A.G. of Hospitals, Bikaner. This pilot study was performed on 200 randomly selected voluntary blood donors (188 men and 12 women, (age range, 18–60 years). A total of 200 donors were typed for D, C, c, E, e, K,k, Jk (a), Jk (b), P1, M, and N, S, s, Fy (a), Fy (b) antigens were typed using antisera from ImmucorInc employing tube methods (Indirect Antiglobulin Test). Antigens frequencies were expressed as percentages.

Results: From the 200 blood donor samples used for extended antigen typing in the Rh system, e antigen was found in 98% donors, followed by D [92%], C [78%], c [71%], and E [30%], with DCe/DCe (R1 R1) as the most common phenotype. K was found to be positive in 2.5% and k=99.5%of donors. Frequencies of M (88%), N (71.5%), S (53.5%) and s (89%), Kidd blood group system antigens (Jka = 79%, Jkb = 64%), (Fya = 84.5%, Fyb =56%), P1=87%were significantly different than other ethnic group.

Conclusion: So knowledge of red cell antigen frequency and phenotype of different regions will helpful in creating donor data bank for preparation of indigenous cell panels and providing antigen negative compatible blood to patients.

2019 May; 13(Suppl 1): S32–S100.

PP 194: Efficient transfusion departmnt: Not only funtion as heart for blood supply but also as strong backbone for successful organ transplant

Background: Organ transplants is the most significant advances in medical practice for saving or extending the lives of people with end-stage organ failure. Over 60 years organ transplantation has been conducted successfully. Indian doctors have opinion that ‘LIVER TRANSPLANT YET TO GAIN ACCEPTANCE “ the high and rising incidence of chronic diseases like diabetes, kidney & liver disease etc will greatly increase the need for organ transplant which is” The best & only cure “from a living or cadaver source.

Materials and Methods: In Pune in 2 ½ years in our hospital 102 transplants were performed for liver Kidney & pancreas. Includes 64 cadaver transplants which are always unplanned cases where the efficiency of transfusion department plays an important role for arranging blood & its components in volume & 38 cases were live transplants.

Results: As per transplant policy minimum 10 units of every components were kept ready, being cadaver transplants the blood bank was always on their toes to meets all requirements for arranging 4080 units. Including all products for above cases only 1412 (34.60%) units were overall utilized as there was no release of entire reserved stock to be maintained at OT & 65.4 % significant percentage of blood could be utilized for other cases. Out of 102 patients 93 (i.e 91%) is our success rate.

Conclusion: With proper camp schedules & voluntary donor support unplanned cadaver Organ transplants could be managed efficiently. Only with proper co-ordination ethical & rational use of blood without wastage was achieved. “TeamSahyadri in coordination with Rotary has made a gunnies world record for “Organ Donation Pledge “from 21,600 people. When team Sahyadri alone could get this achievement, great miracles can be done if we all unite for such noble cause.

2019 May; 13(Suppl 1): S32–S100.

PP 195: Incidence and analysis of 7 years adverse transfusion reaction: A retrospective analysis

Introduction: Safe blood transfusion is the primary need of all the health care delivery system. Though with the advances of transfusion medicine, the incidences of transfusion risk is gradually reduced, but the adverse transfusion reaction (ATR) of non haemolytic type still prevails. The purpose of this study was to estimate the incidence and pattern of transfusion-related adverse events at our centre.

Materials and Methods: The present retrospective observational study was conducted in the Department of Transfusion Medicine, at a multiorgan transplant centre in South India. Adverse Transfusion Reactions reported to the department between April 2011 to April 2018 were included in the study. All the reactions were investigated in detail in the blood bank for the clerical errors, immunohematology workup and classified according to their nature with imputability assessment.

Results: A total of 140 ATR were reported out of 100569 blood components distributed during the study period. After the analysis and workup of the reported reactions, majority of the reactions were observed in males (71%, n=99). Most common symptom presented was Itching/Rashes in 43.57% (n=61) ATR. Allergic reactions (51.42%, n=72), were the most commonly encountered ATR followed by FNHTR (25.71%, n=36). FFP transfusions (0.19%) contributed to the majority of the reactions followed by Red cell transfusion (0.148%). ATR were observed maximum in Liver disease patients (62%) followed by oncology patients (15%).

Conclusion: The overall incidence of ATR in our study is 0.139% which is comparatively low compared to other studies due to well established hemovigilance systems. Adoption of more equipped methods & sensitive technology in various areas of blood banking will help to bring down the unwanted adverse transfusion reactions.

2019 May; 13(Suppl 1): S32–S100.

PP 196: Root cause analysis of incompatible crossmatch at a tertiary care teaching hospital

Background: Blood transfusion is an essential part of therapy for many patients. Although life-saving for many patients, blood transfusion is not without risk. The main goal in transfusion medicine is that transfused blood should be compatible with the patient. The clinical and serologic evaluation, which allows for the transfusion of the most compatible (or “least incompatible”) blood, requires a joint effort between the clinician and the transfusion medicine physician. The transfused Red Blood Cells (RBC's) will have acceptable survival rate and there will be no significant destruction of recipients own RBC's as well as detection of most clinically significant alloantibodies.

Aims: Root cause analysis of incidence & causes of incompatible cross matches in patients by column agglutination method.

Methods: In this prospective study, total of 582296 cross matches were performed over period of last 4 & half years, out of which 867 units were found incompatible by column agglutination method-CAT in polyspecific (IgG + C3d) gel media. A root cause analysis protocol was formulated to resolve incompatibility to ensure safe transfusion to patients.

Results: On the evaluation of 582296 patients’ samples, only 867 units were found to be incompatible (0.14%). The major cause for incompatibility found in patients was autoimmune hemolytic anaemia (32.87%). Other causes of incompatibility were infections (27.44%), multiple transfusions (17.41%), trauma (11.23%), evan's syndrome (4.15%), hemolytic disease of new-born (3.57%), sickle cell anemia (2.99%) and incompatibility due to DAT-Direct Agglutination Test positive Packed Cell Volume (0.34%). Majority of incompatible cross matches in patients were found in females than in males. The most common antibody was found was anti-’M’ and anti-’c’.

Summary: The commonest cause of incompatibility was autoimmune haemolytic anaemia (32.87%). Incompatibility was found more in females (51.44%) than in males. Most incompatible units were found between age <12 (32.87%). Most incompatible units were found in O positive blood group (33.10%). Clerical and technical error has very low incidence. The RCA protocol involves a thorough evaluation of the patient's clinical condition and underlying pathology to identify the cause. A logical stepwise approach will enable provision of safe transfusion to the patient.

2019 May; 13(Suppl 1): S32–S100.

PP 197: A retrospective study on major causes of discarding the blood and its components and to control the wastage from avoidable reasons

Background: Blood transfusion is essential part of modern health care. Human blood has no complete substitute till date. There should be no wastage due to avoidable reasons.

Aim: To find out the major causes for discarding whole blood or components and develop strategy to reduce wastage of blood.

Materials and Methods: A retrospective study of discard of blood & blood components was carried out from July 2017 to June 2018 using records available in blood bank, RNT Medical College, Udaipur. The data were collected from donor record, TTI testing, component preparation & discard records & analysed various reasons for blood bag discard such as seropositivity, hemolysis, clots, less quantity, punctured and expiry.

Results: A total of 21831 blood bags were collected during aforesaid period from which 16512 FFP units and 3228 RDP units were prepared. Total 289 (1.32%) units of WB/PRBC discarded, out of which Major cause was HbsAg (68.85%). Total 204 (1.23%) units of FFP was discarded, out of which major cause was HbsAg (51.96%). Total 330 (10.22%) units of RDP was discarded, out of which major cause was expiry (88.78%). It is seen that wastage of R.D.P. is highest among all blood components. Seropositivity leads to maximum wastage of blood and blood components. Thanks to our FIFO policy, no single unit of PRBC, WB and FFP was discarded due to expiry.

Conclusion: Most common reason for discard of blood was seropositivity. So a simple, non costly, effective and rapid method should be developed for TTIs during screening. Strict FIFO policy should be there. Regular audit will also help to reduce blood wastage. Proper screening of donor and detailed history may also avoid wastage. Bonding between clinician and blood bank staff may also reduce wastage due to improper use, expiry of blood and component specially to reduce R.D.P. wastage.

2019 May; 13(Suppl 1): S32–S100.

PP 198: The perception of haemovigilance among doctors and health care provider: An institutional study

Background: Haemovigilance program of India (HvPI) was initiated in the year 2012 and is a continuous process of data collection and analysis of transfusion-relates adverse reactions in order to investigate their causes and outcomes to preventrecurrence. To improve the existing transfusion services and to check the under reporting transfusion reactions. The present study was done to ascertain the knowledge attitude and practice (KAP) of haemovigilance among doctors and health care providers (HCP). The information thus collected would facilitate corrective and preventive actions to be taken to minimize the potential risk associated with blood collection processing and transfusion to patients.

Aims and Objectives: (a) To assess the knowledge, attitude and practice (KAP) of HCP regarding haemovigilance. (b) To identify the factors which either encourage or discourage transfusion reactions reporting. (c) Generate evidence based recommendation and creates awareness amongst the HCP. (d) Communicate the findings to all key stakeholders.

Methods: It is a cross sectional, pre-validated questionnaire based study carried out among 405 doctors and health care providers in our hospital after obtaining their consent.

Results: The response rate of the results was 98%. 70% of the responders had poor knowledge whereas, 10% of the responders had good knowledge about HvPI. The awareness of reporting of the transfusion reactions was unsatisfactory with only 21% of the responders having the adequate and relevant information.

Conclusions: In order to improve HvPI, it is essential to improveKAP of the HCP. This study will not directly benefit the participants, but their knowledge and practice will safeguard the wellbeing and healthcare of society. The understanding of KAP regarding HvPI among the HCPis the highest standing determinant of their active participation in HvPI implementation. Hence the present study has beendone using the KAP model as a survey tool.

2019 May; 13(Suppl 1): S32–S100.

PP 199: Family history during screening blood donors: Do we really give much importance?

Background: Familial correlations of Hepatitis B and Hepatitis C infection has been published previously in general population. The risk of this infections in blood donors from their families have not been diligently explored.

Aim: Analysis of consecutive donors reactive for Transfusion Transmitted infection (TTI) by fourth generation ELISA.

Materials and Methods: A retrospective study was conducted from January 2016 to July 2018 in a tertiary care hospital, New Delhi. Consecutive donors reactive by ELISA (fourth generation) for HIV, HBV and HCV infections were noted and evaluated.

Results: Out of the 51782 donations 1061 donors were reactive for TTI during our study period. Prevalence of HBV, HIV and HCV infection was 1.32%, 0.21% and 0.51% respectively. Consecutive donors reactive for HBV were 9.20% (63/685), for HCV were 6.0% (16/266) and nil for HIV. In 63 consecutive donors reactive for HBV, 3.80% (26/685) were related to each other, 1.40% (10/685) were not related to each other when donating for the same patient and 3.90% (27/685) were not for the same patient. In 16 consecutive donors reactive for HCV 2.30% (6/266) were related to each other when donating for the same patient and 3.80% (10/266) were donating for different patients. These findings were found statistically significant (p<0.0001).

Conclusion: Among all TTI reactive donors 7.4% (79/1061) were consecutive reactive. The reason for the same may be donor or process related. Donor related reasons may be, virus is transmitted from one of the infective family members. Process related reasons may be cross contamination of the consecutive samples, faulty pipetting etc. In our study 32 reactive donors had close contacts with persons having history of infective disease. Hence, family history for risk factors in donor questionnaire shouldn’t be missed to further reduce TTI.

2019 May; 13(Suppl 1): S32–S100.

PP 200: When variability is the name of the game – The role of monospecific direct antiglobulin test

Background: Direct antiglobulin test (DAT) is a very common immuno-haematological test that is performed. The identification of what is coating the red cells has a great impact on the clinical management. Routine DATs use polyspecific anti-human globulins (AHG) and are able to detect Immunoglobulin-G (IgG) and complement components (c3c, c3d) on red blood cells. However, clearly the in vivo significance of different immunoglobulin isotypes/complement coating, impacts variably. Clinical management of these variations are also different. Monospecific DAT helps define the reason for DAT positivity in most cases.

Aim: The aim of this retrospective study was to describe the profile of Immunoglobulin/complement positivity identified in patients on whom Monospecific DAT was performed during the last six months.

Results: Out of the 45 patients tested, 19 (42.2%) patients showed an isolated IgG positivity. IgG with C3d was present in 11 (24.4%) patients. IgM along with IgG and complements was seen in 3 (6.7%) patients. Isolated complement components (c3c, c3d) positivity was observed in 5 (11.1%) patients. 7 (15.6%) cases were not interpretable as the controls failed.

Discussion and Conclusion: Wide variability of antibodies detected on Monospecific DAT is critical to our understanding the primary disease due to the variability of clinical presentation and different management choices. The in vivo significance of a given degree of RBC sensitization by autoantibodies varies greatly, even among antibodies of the same immunoglobulin class. Corticosteroids which are an initial therapeutic tool in treating patients with autoimmune hemolytic anemia (AIHA) of warm antibody type is less effective for CAS. It is critical that particularly in patients with suspected autoimmune hemolysis, where management can vary greatly based on the type of AIHA, a monospecific DAT be done to help aid diagnosis.

2019 May; 13(Suppl 1): S32–S100.

PP 201: Evaluation of massive blood transfusion protocol practices; single center experience from a tertiary care setup

Background: Concepts regarding the massive transfusion in hemorrhagic shock are constantly changing. This study evaluates massive transfusion protocol (MTP) practices at a tertiary care transfusion center.

Methods: A retrospective review was performed of all the MTP activation episodes. Patient records were reviewed for demographics, indication, blood components transfused, lab parameters and outcome. Blood components (PRBC, FFP, RDP) were given in 1:1:1 ratio. Appropriateness of activation of MTP was assessed on case to case basis. Data management was done using Microsoft Excel Worksheet. Descriptive statistics and paired t test was used for the analysis.

Results: Massive transfusion protocol (MTP) was activated for a total of 105 cases over a period of two and half year (Jan 2015 to June 2017). With a male to female ratio of 1:3. Majority of the requests for massive transfusion were from trauma unit (43%) followed by Obstetric unit (19%). Blood group ‘O’ was the commonest type (41%) among the patients. Prior to the activation of MTP, 14 patients (13%) received 1 to 3 units of Packed Red Blood Cells transfusion. Only one pack (2 PRBC: 2FFP) was used in 56 patients (53%). MTP activation was appropriate in 44% of the cases where patient had actually received massive transfusion. On comparing the hematological parameters between the group of patients with massive blood loss versus the group where MTP was activated in anticipation of massive blood loss, statistically significant difference was with respect to mean hemoglobin (6.7 Vs 10 g/dL) and fibrinogen levels (150 Vs. 188mg/dL). Outcome was favorable in 71 (68%) of the patients.

Conclusion: The present study shows that MTP was activated in anticipation of massive bleed in significant number of cases. Hence we modified the existing protocol incorporating the point of care testing for real time monitoring and to provide individualized transfusion therapy.

2019 May; 13(Suppl 1): S32–S100.

PP 202: A study on role of platelet rich plasma in androgenic alopecia

Background: Androgenetic alopecia is a hereditary, androgen-dependent dermatological disorder more common in men. It is occasionally seen in women. It commonly begins by 20 years of age and affects nearly 50% of men by the age of 50 years. The activated autologous Platelet Rich Plasma (PRP) has growth factors which induce the proliferation of dermal papilla cells. The basic idea behind PRP injection is to deliver high concentrations of growth factors to the scalp, with the hope of stimulating hair regrowth.

Materials and Methods: In our study, 12 young male and 4 female patients suffering with alopecia in the age group of 25-35 years were selected. After taking adequate consent, they were given autologous PRP injections on the affected area of alopecia with insulin syringes in a therapeutic dose of 1.5 million platelets per ml. The injections were given over a period of 3 months at the interval of 2-3 weeks and results were assessed. PRP is generated by taking 30 ml of blood before surgery; this blood is centrifuged at 1000 rpm for 10 minutes (low spin). The plasma is removed and centrifuged at 2000 rpm for 15 minutes (hard spin). The buffy coat is removed and this is PRP.

Results: 3 months after the treatment patient presented with growth in the number of hair, thickness, hair strength and overall alopecia.

Discussion: Androgen-dependent processes are predominantly due to the binding of Dihydrotestosterone (DHT) to the Androgen Receptor Conversion of testosterone to DHT within the dermal papilla plays a central role, while androgen-regulated factors deriving from dermal papilla cells are believed to influence the growth of other components of the hair follicle. The main growth factors involved in the establishment of hair follicle are VEGF, epidermal growth factor (EGF), insulin 1-like growth factor, and fibroblast growth factor.

Articles from Asian Journal of Transfusion Science are provided here courtesy of Wolters Kluwer -- Medknow Publications