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Background and Objectives: Correlation has been shown between hepatitis B surface antigen (HBsAg) and hepatitis B virus (HBV) levels in different patient categories. However, this has not been studied for donors. The aim of this study was to study the relationship between HBV DNA levels and E ratio obtained for HBsAg testing by chemiluminescence immunoassay (CLIA) performed on donor samples. Methods: A prospective, observational study was conducted in the department of transfusion medicine at a large tertiary health-care setup in North India from 2017 to 2021. Serological testing for HBV included HBsAg testing by CLIA. All reactive samples were tested in duplicate by electro CLIA (eCLIA). All samples found to be reactive in all three tests were then tested by a rapid diagnostic test and quantitative polymerase chain reaction. Results: During the study duration, 251 donors were found to be reactive for HBsAg by CLIA and eCLIA in screening. Of these 251, HBV DNA could not be detected in 85 (33.86%) samples, whereas in rest 166 (66.14%) samples, HBV DNA was detected. Mean E ratio for CLIA for all 251 samples was 5177.45, whereas mean viral copies detected were 19645858.66. The correlation for all 251 samples was found to be weak negative. The correlation for sero-reactive samples with the presence of HBV DNA was also found to be weak negative. Conclusion: In conclusion, we found significant but weak correlation between HBsAg results obtained by CLIA and HBV DNA levels for blood donors. The measurement of HBV DNA level is necessary for all apparently healthy donors coming for blood donation and testing reactive for HBsAg in screening assays. [ABSTRACT FROM AUTHOR]

Despite knowing benefits of extended phenotyping, a vast majority feel that phenotype matched units add to the cost of blood banking. The purpose of this study was to discuss advantages and disadvantages of performing Rh Kell phenotyping in Indian scenario. This was a prospective, observational study conducted at a tertiary healthcare center between July 2014 and February 2020. All consecutive whole blood donors and all consecutive patients whose samples were sent for Rh-Kell phenotyping were included for calculating antigen, phenotype and gene frequencies. For rate of alloimmunization in patients transfused with phenotype matched units, all patients who were given Rh-Kell phenotype matched transfusions were included in the prophylactic antigen matched (PAM) category and those who were given random units were included in the non-PAM category. A total of 37,588 donors and 258 patients were included in the study for calculation of antigen, phenotype and gene frequencies. Percentage similarity of phenotypes between patient and donor populations was 33.8%. For rate of alloimmunization, results of a total of 31,991 patient samples revealed 0.94% prevalence of unexpected antibodies; highest against the Rh system. Three patients in the non-PAM category and one in the PAM category were alloimmunized during follow-up. Significant clinical and laboratory impact of phenotyping was observed in terms of reduced turnaround time and consumption of resources. Rh-Kell phenotyping of donors can prevent alloimmunization, reduce cost burden on the patient and the laboratory and help the laboratory personnel in smooth routine testing. [ABSTRACT FROM AUTHOR]

BACKGROUND: Therapeutic plasma exchange (TPE) has been advocated as an adjunct to steroids and cytotoxic drugs in treating patients suffering from vasculitis and presenting with active disease, but we still have insufficient evidence on its effectiveness in improving the clinical response, especially in India. This study was planned to study the clinical outcome in severe vasculitic presentations treated with TPE as an adjunctive therapy. MATERIALS AND METHODS: A retrospective analysis of TPE procedures performed from July 2013 to July 2017 in the department of transfusion medicine at a large tertiary care hospital was done. All consecutive patients admitted with new diagnosis of systemic vasculitis presenting with active disease and severe presentations such as advanced renal failure or severe respiratory abnormalities or life-threatening vasculitis affecting the gastrointestinal tract, neurological and musculoskeletal system; who needed TPE for removal of preformed antibodies, were included in the study. RESULTS: There were a total of 31 patients in whom TPE was performed for severe systemic vasculitis; 26 adults and five pediatric. Six patients tested positive for perinuclear fluorescence, 13 for cytoplasmic fluorescence (cANCA), two for atypical antineutrophil cytoplasmic autoantibody, seven for anti-glomerular basement membrane antibodies, two for antinuclear antibodies (ANA), and one patient tested positive for ANA as well as cANCA before the augmentation of TPE. Out of 31, seven patients showed no clinical improvement and succumbed to the disease. At the end of desired number of procedures, 19 tested negative and five tested weak positive for their respective antibodies. CONCLUSION: Favorable clinical outcomes were observed with TPE in patients with antibody-positive systemic vasculitis. [ABSTRACT FROM AUTHOR]

BACKGROUND AND AIMS: When determining ABO antibody titers, immunoglobulin G (IgG) antibodies can be masked by immunoglobulin M (IgM) antibodies. Hence, the measurement of actual concentration of IgG requires methods like heat inactivation (HI) of plasma. This study was aimed at determining the effects of HI on IgM and IgG titers performed by conventional tube technique (CTT) and column agglutination technique (CAT). MATERIALS AND METHODS: This was a prospective, observational study conducted from October 2019 to March 2020. All consecutive A, B, and O group donors who gave consent for participation were included. All samples were consecutively tested by CTT and CAT, before and after HI (pCTT, pCAT). RESULTS: A total of 300 donors were included. IgG titers were found to be more than IgM titers. For group O, IgG titer results were higher for both anti-A and anti-B compared to group A and B. For group A, B, and O, pretreatment results were higher than posttreatment IgG titer results. Median anti-A titers were similar to median anti-B titers across all categories. Median IgM and IgG titers were higher for group O individuals than nongroup O individuals. There was reduction in IgG and IgM titers after HI of plasma. One log reduction in median titers was observed when ABO titers were performed by CAT and CTT. CONCLUSION: There is one log difference between median antibody titers estimated using heat inactivated and nonheat inactivated plasma. The use of HI for ABO isoagglutinin titer estimation can be considered in low resource settings. [ABSTRACT FROM AUTHOR]

Background and aims: In order to determine actual concentration of clinically significant IgG antibodies, there is a need to inactivate IgM antibodies, which can be performed by DTT or heat. The aim of the present study was to compare the effect of DTT treatment and heat inactivation (HI) on ABO isoagglutinin titres performed by column agglutination technology (CAT) and conventional tube technique (CTT). Materials and methods: This was a prospective, observational study conducted from October 2018 to March 2020. All consecutive O group donors were included. Serum from each donor was treated with DTT, and heat and tests were performed by CTT and CAT before and after treatment. Results: A total of 2005 donors were included. 401 (20%) samples had inter‐observer variation by CTT, and 69 (3·4%) had inter‐observer variation by CAT. Median anti‐A and anti‐B titre results were higher with CAT as compared to CTT. Median anti‐A and anti‐B IgG titre results showed a one‐ to twofold difference between CTT and CAT with higher titres reported by CAT. Median IgM titres decreased two‐ to threefold after DTT treatment, whereas onefold decrease was confirmed after HI. Concordance was found to be poor. However, correlation was found to be strong. Conclusion: There is overestimation of ABO IgG antibody titres and hence, estimation of ABO IgG titres after inactivation of IgM antibodies is strongly recommended. DTT treatment was found to be superior to HI in terms of efficiency in elimination of IgM activity. [ABSTRACT FROM AUTHOR]

Background and aims: To determine actual concentration of clinically significant IgG antibodies, there is need to inactivate IgM antibodies. Aim of this study was to study effect of dithiothreitol (DTT) on ABO isoagglutinin titres performed by column agglutination technology (CAT) and conventional tube technique (CTT) and to compare results obtained by DTT treatment. Materials and methods: This was a prospective, observational study conducted from October 2019 to March 2020. All consecutive A, B and O group donors were included. All samples were consecutively tested by CTT and CAT, before and after DTT treatment (pCTT, pCAT). Results: A total of 300 donors were included, 100 each from group A, B and O. IgG titres were found to be more than IgM titres. For group O, IgG titre results were higher for both anti‐A and anti‐B compared with group A and B. Reduction in IgG titres was observed with DTT. With the use of DTT, one log reduction in group O IgG titres was observed and three log reduction in group A/B IgG titres was observed. When comparing anti‐A and anti‐B IgG results for group O individuals, results of CTT and pCAT were not found to be significant. IgG measurement of anti‐A and anti‐B of group O individuals showed strong to very strong correlation between CTT and CAT, before and after DTT treatment. Conclusion: There is significant difference between antibody titres estimated using DTT‐treated and DTT‐untreated plasma. Use of DTT for ABO isoagglutinin titre estimation is recommended. [ABSTRACT FROM AUTHOR]

Objectives: The aim of this study was to analyse blood requisition forms sent by clinicians in a tertiary care hospital to the transfusion service to ascertain their completeness and correctness. A secondary objective was to study the effect of continuing medical education (CME) in a hospital setting on clinician's behaviour regarding the importance of details that ought to be mentioned on blood requisition forms. Background: Transfusion audits are useful tools in the evaluation and education of those requesting blood components. Methods/Materials: This was a prospective, observational study conducted in the department of Transfusion Medicine at a tertiary‐level healthcare centre from June 2019 to December 2019. The study was divided into two phases: pre‐CME (P1) and post‐CME (P2). In both phases, an audit for assessing completeness and correctness of blood requisition forms, which were divided into four sections, was performed. A scoring system was devised to compare both phases. Results: In the P1 phase, 45.77% of the blood requisition form entries were complete and correct; 23.45% of incomplete entries were generated by emergency and trauma. In the P2 phase, 76.75% of the blood requisition form entries were complete and correct; 35.09% of the incomplete entries were generated by obstetrics and gynaecology. Complete and correct entries increased from 45.7% (P1) to 76.75% (P2). Scores of P1 were found to be lower than scores of P2 for all four sections. Cumulative mean score for P1 (20687) was found to be significantly lower than the mean score for P2 (30870). Conclusion: Audit and CME regarding different aspects of transfusion medicine practices play a major role in the improvement of transfusion practices in hospitals. [ABSTRACT FROM AUTHOR]

Background and aims Antibody titration by the manual method has certain limitations. The aim of this study was to compare the manual conventional test tube technique (CTT) with semi-automated, column agglutination technique (CAT) and fully automated haemagglutination/solid-phase red cell adherence (HA/SPRCA) technology in measuring antibody titres of anti-A and anti-B isoagglutinins of blood group O blood donors. The median IgM and IgG antibody titres with 1+, 2+ and 3+ end-points were determined for each technology, and the agreement and correlation between results of the 3 technologies were evaluated. Materials and methods This was a prospective, observational study conducted from October 2018 to March 2020. All consecutive O group blood donors, who donated blood at the Department of Transfusion Medicine at a tertiary healthcare centre, were included in the study. All samples were consecutively tested by CTT, CAT and HA/SPRCA, and the results were recorded and compared. Results A total of 2005 donors were included. Inter-observer variation was observed in 295 (14.71%) samples by the CTT and in 51 (2.54%) by the CAT. There was 43.4% and 40.3% agreement of anti-A IgM and IgG titres between CAT (2+ strength) and CTT (1+ strength), respectively. The agreement for anti-B IgM and IgG titres between CAT (2+ strength) and CTT (1+ strength) was 43.6% and 32%, respectively. While a strong correlation was found between CTT (1+ strength) and CAT (1+, 2+ strengths) for both IgM and IgG measurements of anti-A and anti-B isoagglutinins, the correlation between CTT (1+ strength) and HA/SPRCA (1+, 2+ strengths) was found to be weak. The median IgM titres for anti-A and anti-B were lower than the median IgG titres. Conclusion The antibody titration levels were higher with CAT than with CTT, and those of HA/SPRCA were lower than those of CTT. Transfusion services need to decide which method would provide higher benefit to their patient population. [ABSTRACT FROM AUTHOR]

Anti-G antibody mimics the reactivity pattern of coexistent anti-D and anti-C. Differentiating between the two is significant in antenatal females where the decision to administer RhD prophylaxis is based on the presence or absence of anti-D antibody. The aim of reporting this serological challenge is to emphasize the need for phenotyping red cells for sourcing appropriate in house red cell reagents and to help transfusion services sharpen problem-solving skills. A 26-year-old pregnant female with a complicated obstetric history and a positive indirect antiglobulin test presented to the hospital for antenatal assessment at 24 weeks. A positive antibody screen warranted identification of the implicating antibodies. Since identification was suggestive of multiple alloantibodies whose specificities could not be confirmed, step-wise sequential adsorption and elution was required. Anti-D, anti-C, and anti-E antibodies were identified in patient plasma with titers of 1024, 4, and 32, respectively. The absence of anti-G was also confirmed. Multiple alloantibodies can pose a challenge to transfusion services. However, with the help of select cells, phenotyping, adsorption elution studies, and phenotyped donor units; solving complex serological cases can be accomplished. [ABSTRACT FROM AUTHOR]

Objectives: The primary aim was to assess the knowledge, attitude and practices (KAP) of employees (medical, paramedical and other hospital staff) towards blood donation and transfusion practices in a tertiary care hospital. A secondary objective was to assess and interpret the effect of an educational module on improvement in the KAP of employees in a hospital setting. Background: For satisfactory practices, it is essential to initiate KAP studies. Methods/materials: This was a prospective, observational study conducted among hospital staff (clinical and non‐clinical) from January to December 2019. The study was divided into two phases: pre‐educational module (P1) and post‐educational module (P2). In both phases, a questionnaire was distributed. Knowledge was assessed by 30 questions, attitude by 20 questions and practice by 30 questions. If any individual had unsatisfactory scores in both the P1 and P2 phases (scores <40; 50%), they had to participate in a mandatory certificate course. Results: A total of 180 individuals participated in the P1 and 172 participated in the P2 phase. Mean score for practice (0.471) was better than that for attitude (0.447) and knowledge (0.43). Factors associated with good scores were younger age group, more than 5 years of employment and clinical field of study. The total score for KAP increased, and a statistically significant difference (P‐value <.05) was observed between P1 and P2 scores. Of 172 participants, 27 were asked to attend the 2‐week certificate course due to unsatisfactory scores (score < 40) in both P1 and P2 phases. These 27 participants required attending this certificate course a mean of 1.67 ± 0.83 times to obtain satisfactory scores. Conclusion: Educational intervention is an important tool for improving KAP among not only physicians but hospital staff as well. [ABSTRACT FROM AUTHOR]

BACKGROUND: Erycard 2.0 is a "point-of-care" device that is primarily being used for patient blood grouping before transfusion. MATERIALS AND METHODS: Erycard 2.0 was compared with conventional slide technology for accuracy and time taken for ABO and Rh forward grouping result with column agglutination technology (CAT) being the gold standard. Erycard 2.0 as a device was also evaluated for its stability under different storage conditions and stability of result till 48 h. In addition, grouping of hemolyzed samples was also tested with Erycard 2.0. Ease of use of Erycard 2.0 was evaluated with a survey among paramedical staff. RESULTS: Erycard 2.0 demonstrated 100% concordance with CAT as compared with slide technique (98.9%). Mean time taken per test by Erycard 2.0 and slide technique was 5.13 min and 1.7 min, respectively. After pretesting storage under different temperature and humidity conditions, Erycard 2.0 did not show any deviation from the result. The result did not change even after 48 h of testing and storage under room temperature. 100% concordance was recorded between pre- and post-hemolyzed blood grouping. Ease of use survey revealed that Erycard 2.0 was more acceptable to paramedical staff for its simplicity, objectivity, and performance than conventional slide technique. CONCLUSION: Erycard 2.0 can be used as "point-of-care" device for blood donor screening for ABO and Rh blood group and can possibly replace conventional slide technique. [ABSTRACT FROM AUTHOR]

Immunohematology reference laboratory (IRL) is defined as a fully equipped advanced laboratory that receives samples from various parts of the country for workup. This is a specialized center, where other centers experiencing difficulty in resolving immunohematological tests, refer samples for further workup and resolution. To complete the workup process, ABO RhD typing, antibody screening, identification (ID), antigen phenotyping, and direct antiglobulin tests are performed. In our retrospective observational study 528 cases, worked up from October 2019 to March 2022 were included. The prevalence of alloimmunization found in this evaluation was 68.1%. Among the different alloantibodies, anti‑D was most common followed by anti‑E and anti‑M. In our study, most of the autoantibodies were of Immunoglobulin G (IgG) class (79.31%). Out of the 528 samples, ABO discrepancy was found in 48 samples (9%). Of these, none belonged to Group I or Group III discrepancy types. Group II discrepancies were noted in 28 cases (58.3%) and Group IV discrepancies were found in 20 cases (41.6%). Our study clearly indicates that there is an important role of IRL which is not only limited to ID of alloantibodies or resolution of ABO discrepancies but also supports all the hospitals and transfusion centers in the region by providing phenotype-matched or compatible units and advice for safe transfusion practices. [ABSTRACT FROM AUTHOR]

Preconditioning using different protocols has been tested to prevent antibody mediated rejection (ABMR) individually for ABO and HLA incompatibility. However, simultaneous presence of both barriers is still less explored. The aim of this study was to report outcomes of institutional desensitization protocol in renal transplant recipients with simultaneous ABO and HLA incompatibility. This was a retrospective study conducted from October 2015 to December 2018. All patients with a clinical diagnosis of dialysis dependent chronic kidney disease (CKD), who were prospective coexistent HLA and ABO incompatible renal transplant recipients were included in the study. Patients were followed up and graft function and patient survival was assessed at 1 y from the date of transplant. Median and mode baseline anti-A titers were 64, while median and mode baseline anti-B titers were 256. All recipients were discharged by tenth postoperative day. None of the patients had any bleeding complications. Post transplant infection rate was found to be 20 %. A total of 54 therapeutic plasma exchange (TPE) procedures were performed before transplant and 8 were performed after transplant. Graft survival and patient survival was 100 % at 3, 6, 9, and 12 months. Range and mean follow-up period was 15–42 months and 23 months respectively. Mean glomerular filtration rate (GFR) at 1 y using the CKD-EPI equation was 85.25 ± 13.76 mL/min. Biopsy proven ABMR was observed in one case only which was managed with TPE and immunosuppression. Simultaneous ABO and HLA incompatibility in renal transplant recipients can be managed successfully with adequate preconditioning and careful monitoring. [ABSTRACT FROM AUTHOR]

Anti-Cw antibody is an immunoglobulin against the red cell antigen Cw which is a low frequency red cell antigen that belongs to the Rh antigen system. It is a clinically significant antibody and may cause haemolysis on exposure to antigen positive red cells. Due to its low frequency, it is not included in routine antibody screening panels. A 32 years healthy male donor with no history of transfusion donated whole blood at the department of Transfusion Medicine & Blood Centre of our institute. As a part of routine pre-transfusion testing, the donor's samples were subjected to automated blood grouping and screening for unexpected red cell antibodies using 3 cells panel on solid-phase red-cell adherence (SPRCA) (Galileo Neo, Immucor, Norcross, USA). The antibody screening came out to be positive with a reaction in cell I of the antibody screening panel. Further the antibody was identified as anti Cw in using 16 cells panel, select cells and phenotyping. In the present case, the anti-Cw antibody was found to be reactive at 37 °C and AHG phase which could lead to haemolytic transfusion reaction. The fact that the male donor had no history of transfusion or transplant led us to the conclusion that it was a naturally occurring, but a clinically significant antibody. This case highlights the importance of performing an antibody screening for healthy donors as well and urges transfusion services to procure screening cells which incorporate Cw positive cells. [ABSTRACT FROM AUTHOR]

Background and Objectives: Audits identify areas of problems in immunohematology workup and transfusion practice which can be corrected by the education of medical staff, formulation of guidelines, and algorithms. An audit is a series of simple, direct questions, which when answered and reviewed, tell us whether the laboratory is performing its procedures, activities, and policies correctly. The aim of this study was to analyze immunohematology request forms sent from red cell serology laboratories to our reference testing center (RTC) to ascertain their completeness. Materials and Methods: This was a retrospective, observational study conducted in the department of transfusion medicine at a tertiary-level health-care center from November 2019 to June 2021. We ascertained whether the immunohematology workup forms details were complete, absent, or incomplete. Results: The study was conducted over a period of 20 months, with 264 forms being audited. In this overall total number, almost one-third of 1894 (34.16%) of the immunohematology workup requisition form details were complete and 716 (12.91%) of these entries were incomplete. However, almost half of the overall number of 2934 (52.92%) of the requisitions were absent. Conclusion: From our study, we concluded that an audit of all the RTC requisitions at the point of receiving can be an important tool to detect requisition errors and results of RTC workup. We received a sample for RTC advanced immunohematological testing from all over India. There is a need for the organization of CME and training of staff to increase compliance regarding sending appropriate RTC advanced immunohematological requisitions and samples. [ABSTRACT FROM AUTHOR]

ABO subgroups or weaker variants of A or B are group A or B subjects whose erythrocytes give a weak or negative reaction serologically with anti-A or Anti – B antisera respectively. Occurrence of these subgroups may lead to an ABO discrepancy which often puts transfusion services in a quandary. ABO subgroups which present as ABO discrepancies can be missed if reverse grouping is not performed. This study was planned to estimate the prevalence of different subgroups which can present as an ABO discrepancy in Indian population, and provide an insight to transfusion services for identification of subgroups serologically. A cross-sectional, analytical study was performed at a tertiary healthcare based blood bank on whole blood donors and patients from January 2017 to July 2018. All suspected type II and Type IV (with Anti-A1) ABO discrepant samples were projected to an algorithmic testing process, to confirm discrepancy and then narrow down to the probable subgroup. A total of 33 subgroup discrepancies; 26 of A group and 7 of B group were identified out of 73,380 patient and 35,279 donor samples tested for blood grouping. Following the algorithm, the overall prevalence of weak subgroups which can present as an ABO discrepancy was found to be 1 in 3293 or 0.03% in our population by serological testing. Out of the discrepancies caused by subgroups, the prevalence of subgroups of A were 0.0101%, 0.0018%, 0.0009%, 0.0027%, 0.0027% and 0.0018% for A2 with anti-A1, A3, Aend, Ax, Am and Ael respectively while those of B were 0.009%, 0.0009%, 0.0009% and 0.009% for B3, Bx, Bm and Bel respectively. Algorithmic approach for resolution of ABO discrepancies caused by subgroups helps in identifying the subgroup which is important because these individuals may be mistyped as group O individuals. [ABSTRACT FROM AUTHOR]

Achievement of complete donor chimerism after an allogeneic hematopoietic stem cell transplant is necessary for elimination of underlying malignant disease. A decline in donor chimerism may herald an impending relapse and therefore, early recognition and intervention plays an important role in such cases. A 32 year old male patient diagnosed as a case of Philadelphia positive mixed phenotypic acute leukaemia underwent peripheral blood hematopoietic stem cell transplant (HSCT) with his sibling as donor. During follow-up, a fall in donor chimerism was observed from 91.86% on day +37 to 88.83% on day +57 and 85.34% on day +77. Donor Lymphocyte Infusion (DLI) was harvested via apheresis. A dose of 1 × 106 per kg was infused and the rest was cryopreserved in aliquots of escalating doses. On day +102, he presented with biopsy proven acute mucocutaneous GVHD grade 2 which was managed conservatively and donor chimerism of 57.99%. On day +126, a repeat donor chimerism was performed which showed 100% chimerism. He continues to do well at day +161. Timely use of DLI can improve donor chimerism in patients with Philadelphia positive acute leukaemia who tend to relapse after HSCT. • Philadelphia positive leukaemia • Donor lymphocyte infusion • Mixed chimerism • Graft versus Host Disease (GVHD) [ABSTRACT FROM AUTHOR]

• 6678 donor blood samples were phenotyped to look for unusual phenotypes in Indian population. • Indigenous antibody screening panel was created with the help of phenotyped voluntary donor registry. • A rare donor with D - - phenotype was registered with the IRDP, Bristol, UK. A clinically significant red cell alloantibody is capable of accelerated destruction of red cells bearing the corresponding antigen. Knowledge of prevalence of these antigens is necessary for performing day to day work and for research in immunohematology. The primary aim of this study was to find the prevalence of 18 clinically significant blood group antigens in blood donors. Secondary objectives were to motivate and create a database of accessible, volunteer O blood group donors and to register rare donors with existing registries. A cross-sectional observational study was conducted in the department of Transfusion Medicine at a large tertiary care hospital in India from October 2016 to May 2018 with a planned sample size of 4800. Study population included healthy blood donors of either gender coming for blood donation to the blood centre. A total of 6678 samples were included in the study. First time donors were 21.41 % while 78.59 % were repeat donors. Voluntary donors constituted 15.81 % while replacement donors were 84.19 %. Male donors were 89.82 % while female donors were 10.18 %. The antigen, phenotype and gene frequencies were calculated. An extended phenotyped voluntary donor database was created and four rare donors were identified. One of these rare donors was registered with the International Rare Donor Panel (IRDP) and rest were registered in a local registry. This study might help enhance the confidence of blood banks in finding appropriate units for patients with unexpected antibodies or with rare phenotypes. It also paves a way for registering rare donors locally and internationally. [ABSTRACT FROM AUTHOR]

Background and Objectives: Measurement of actual concentration of IgG requires methods like heat inactivation (HI) of serum. This study was aimed at comparing of heat-treated ABO titers performed by conventional test tube technique (CTT) and column agglutination technique (CAT) with hemagglutination (HA)/solid-phase red cell adherence (SPRCA). Methods: This was a prospective, observational study conducted between October 2019 and March 2020. All consecutive A, B, and O group donors who gave consent for participation in the project were included. All samples were tested by CTT and CAT before and after HI (post DTT treatment performed by CTT [pCTT] and post DTT treatment performed by CAT [pCAT]) and with HA/SPRCA. Results: A total of 300 donors were included: 100 each with A, B, and O blood groups. For group O, IgG titers were higher than IgM titers among group O, whereas that among group A and group B, IgM titer was found to be higher than IgG titer by both HA/SPRCA and pCTT. The titer values among all the three blood groups were highest when the test was performed by pCAT, though the results of pCTT and SPRCA, pCTT, and pCAT were significant in group O donors. For groups A and B, results of pCTT and pCAT, pCAT, and HA/SPRCA were found to be statistically significant. Conclusion: Results obtained by HA/SPRCA were closer to pCTT, with respect to time consumption, expertise required, and lack of inter-observer variation. Titers obtained by pCAT were higher in comparison to pCTT obviously due to its relative higher sensitivity. Hence, these methods cannot be used interchangeably. [ABSTRACT FROM AUTHOR]

ABO incompatible single donor platelet concentrates (SDPC) have a concern about unsatisfactory increments as well as possibility of hemolytic transfusion reaction. But from Indian population no study has commented on the clinical and laboratory outcome of ABO mismatched platelet transfusion. The aim of study was to compare transfusion outcomes in ABO identical versus ABO non-identical single donor platelet concentrates. In this prospective observational study, 400 SDPC transfusions among different patients were included. In group A (n = 200), ABO identical SDPC transfusions and in group B (n = 200) ABO non-identical SDPC transfusions were added. Corrective count increment (CCI), absolute count increment (ACI), percent platelet recovery (PPR) were calculated and incidents of hemolytic transfusion reactions were noted. In group A mean ± SD of ACI, CCI and PPR were as 30.78 ± 12.51, 15.10 ± 6.677, 39,948.9 ± 20,099.392. In group B, mean ± SD of ACI, CCI and PPR were – 25.4 ± 15.65, 12.509 ± 5.906, 33,559.2 ± 22,150.304. And when CCI, ACI, PPR were compared with group A and group B, statistically significant differences were noted (P < 0.05). There was statistically significant difference in CCI, ACI and PPR in oncology patients and other prophylactic recipients except patients with dengue and other infectious disease. But there was no hemolytic transfusion reaction noted in any group. Our study clearly establish the potential benefits of ABO-identical PLT transfusion. It also points out that in emergency conditions or when there is a paucity in inventory, ABO non-identical SDPC transfusion may be lifesaving and clinically significant. [ABSTRACT FROM AUTHOR]

Introduction: Cell-based complement-dependent cytotoxicity crossmatch (CDC-XM) and solid phase assays were introduced for assessing HLA antibodies. However, the complexity of data from cell-based and solid phase assays have led to potential confusion about how to use the results for clinical decision making. Aim: Aim of this study was to compare results of cell-based assay and solid phase assay, to evaluate the usefulness of L-XM for pretransplant detection of HLA class I and II donor-specific IgG antibodies, correlate the mean fluorescence intensity (MFI) values of class I and class II L-XM assay and with CDC-XM and L-PRA (panel reactive antibodies) results. Methods: In this retrospective study, 380 prospective renal transplant recipients were tested for the presence of HLA antibodies by CDC-XM, IgG-L-XM, IgG-L-PRA & L-SAB screening with their corresponding donor. Results: Fifty-one recipients (13.42%) had a positive CDC-XM. L-XM was positive in 125 recipients (32.89%); class I-L-XM was positive in 46 (36.80%) cases, and class II-L-XM was positive in 58 (46.4%) cases and 21 (16.8%) samples were positive for class I and class II. High background was present in 22 (5.87%) samples, the results of which were confirmed by retesting or by correlation with L-PRA and L-SAB assays. Conclusion: The introduction of more sensitive approaches for the detection of anti-HLA-IgG-antibodies, such as L-XM and L-PRA assay, has allowed the identification of anti-HLA-antibodies in recipient serum which is not usually identified by CDC-XM alone. However, L-XM has some limitations; they can be overcome if we combine this assay with L-PRA. [ABSTRACT FROM AUTHOR]

ABO-incompatible transplantations allow patients to receive timely transplants. Isoagglutinin titration to ascertain levels of incompatible antibodies in the recipient is important in determining patient selection and transplant survivability. To find out the prevalent trends in India, the largest, first of its kind survey was carried out among the transplant centers regarding their practices in isoagglutinin titration. The survey was drafted by a working group of Transfusion and Transplant Immunology specialists from six different centers. Data was obtained via the use of an online questionnaire. Results were categorized into four categories, Hospital information, Titration methodology, Role of transfusion specialists and cut-off titers. Most centers had a well-established solid-organ transplant program with considerable number of ABO-incompatible transplantations. Most centers performed isoagglutinin titration in Transfusion Medicine department. Column Agglutination Technique (CAT) was the most common method, using EDTA blood samples and freshly-prepared in-house pooled cells. Most centers had a turn-around time of less than 12 h. While the policy for ascertaining baseline and threshold titers is well-defined in ABO-incompatible renal transplants, variations from center to center still exist for ABO-incompatible liver transplants. Most centers required a Transfusion Medicine consultation for the patients before such transplants. With increasing ABO-incompatible kidney and liver transplants across the country, the role of Transfusion medicine specialists has become vital in pre-conditioning regimes enabling the viability and success of such transplants. This was a unique survey that provided a snapshot of current trends and practices of isoagglutinin titration for ABO-incompatible transplants in India. • This was a survey undertaken among the Transplant centers across the country performing ABO incompatible transplants • Such transplants are being performed not only at larger tertiary centers in metro cities, but also in smaller hospitals across the country • The protocols for kidney transplants in regards to baseline and threshold titers appear better defined as compared to those for liver transplants • Underscores the growing importance of Transfusion Medicine as an integral part of such complex procedures [ABSTRACT FROM AUTHOR]

There are several reports in medical literature about Red Cell Exchange (RCE) being routinely performed pre-operatively in sickle cell disease patients to provide immediate decrease in HbS concentration and prevent post-operative complications. We would like to present one such case of SCD who also had multiple allo-antibodies and had to undergo hemi-arthroplasty for avascular necrosis of head femur. Grouping and antibody screening was performed using column agglutination technique. 3-cell and 11- cell panel were used for antibody screening and identification, respectively. Automated RBC exchange was performed on apheresis machine Com.Tec using the standard PL1 kit (Fresenius Kabi, Germany). Multiple (anti-c, E) allo-antibodies were identified and successful pre-operative RCE was done with corresponding antigen-negative AHG compatible RBC units. Single RCE procedure reduced HbS concentration from 65% to 25%. The patient underwent uneventful hemi-arthroplasty and was discharged on post-operative day-7. Patient is on regular follow-up and continues to do well two months after the day of surgery. This is possibly the first case report from India, which illustrates successful automated RCE in a SCD patient with alloimmunization. [ABSTRACT FROM AUTHOR]

Several autoimmune disorders have been reported to be related with COVID infection. In continuation to these autoimmune phenomenon, autoimmune hemolytic anaemia (AIHA) also has been noted in COVID infected patients. The aim of the study was to find out the prevalence of red cell alloimmunization, ABO discrepancy and positive direct antiglobulin test (DAT) results in COVID infected patients hospitalised in a tertiary care centre in North India. This was a retrospective observational study done from July 2020 to June 2021. All symptomatic patients admitted to ICU tested positive for SARS CoV-2 whose blood samples were received in the immunohematology laboratory of department of Transfusion Medicine for determination of blood group and issue of packed red cells, and found to have positive antibody screen, blood group discrepancy and positive DAT results, were included in the study. A total of 10,568 tests were run, out of which 4437 were for determination of blood group, 5842 were for antibody screen and 289 were for direct antiglobulin test. Included in this study were 146 patients who either had blood group discrepancy, or had a positive antibody screen or had a positive DAT. Out of 115 positive antibody screen, 66 patients had only alloantibodies, 44 patients had only autoantibodies while only 5 patients had both auto as well as alloantibodies. Total number of positive DAT cases was 50 (50/289 = 17.3 %). There were 26 ABO discrepancies (26/4437 =0.58 %) found. Our results also indicate that there is rise in rate of alloimmunization and DAT positivity among COVID patients. [ABSTRACT FROM AUTHOR]

61-year old male patient was admitted to the hospital with clinical picture of hemolytic anemia with hemoglobinuria. Patient was suspected to have Infectious Mononucleosis (IM) with Auto Immune Hemolytic Anemia (AIHA). DAT was positive with anti-C3d specificity. Donath Landsteiner (DL) test was positive; confirming Paroxysmal Cold Hemoglobinuria (PCH). The final diagnosis was IM with PCH. Patient was managed conservatively and discharged after seven days. DL test specifically detects a biphasic autoantibody (IgG type) that binds to RBCs at cold temperatures, and fixes complement on the RBC membrane. However RBCs are only lysed upon warming to 37C when complement cascade proceeds to completion. [ABSTRACT FROM AUTHOR]

• Sequential serological testing strategy increases diagnostic accuracy. • Sequential testing strategy, previously proven for anti-HIV, can also be applied to HBsAg. • PPV of Strategy III (ECLIA + RDT + ELFA) > Strategy IIB(ECLIA + RDT) > Strategy IIA(ECLIA + ELFA) > Strategy I(ECLIA). • Strategy IIB(ECLIA+RDT) was found to be the best strategy owing to diagnostic accuracy and cost effectiveness. The diagnostic accuracy of any serological test for detection of HBsAg is not 100%. We hypothesized that the sequential testing strategy proven for anti-HIV laboratory diagnosis should also apply to other infectious disease markers like HBsAg. Therefore, we evaluated the diagnostic accuracy of these strategies, I (single test), II (two tests in sequence), III (three tests in sequence) for diagnosis in patients and blood donors and compared it to the confirmatory test for HBsAg (Neutralization Test). Samples were initially tested for HBsAg by A1- Enhanced Chemiluminescent Immuno Assay (ECLIA). Initial reactive (aliquoted donor/patient) samples were reflexly tested by A2- Enzyme Linked Fluorescence Assay (ELFA) and A3- Immuno Chromatography Assay (ICA) assays. Confirmatory neutralization assay was performed on all initial reactive samples. Four strategies (I, II A, II B, and III) that were used in this analysis were; I = A1, IIA = A1 + A2, IIB = A1 + A3, and III = A1 + A2 + A3. The results of all four strategies were compared to Gold Standard (Neutralization Test). A total of, 112, 011 blood samples (75,111 patient samples and 36,900 whole blood donor samples) were initially tested for HBsAg by A-1 (CLIA). Amongst the tested samples, 1,296, 1,188, 1,078, 1,074 samples were found to be reactive by strategy I, IIA, IIB, III respectively. We observed that the PPV (Positive Predictive Value) of Strategy III > Strategy IIB > Strategy IIA > Strategy I. Sequential serological testing strategy comprising of initial sensitive test followed by more specific test increases the diagnostic accuracy of test report as compared to a single test. [ABSTRACT FROM AUTHOR]