Retrospective analysis of forward and reverse ABO typing discrepancies among patients and blood donors in a tertiary care hospital.

OBJECTIVE: The aim of our study was to determine the incidence and causes of ABO typing discrepancies among patients and blood donors at our centre. BACKGROUND: An accurate interpretation of the ABO blood group of an individual is of utmost importance to ensure patient safety and good transfusion practices. METHODS: A retrospective observational study was carried out in the Department of Transfusion Medicine in our hospital from March 2013 to December 2015. Records of all patient and blood donor samples were retrieved and analysed for ABO typing discrepancies. RESULTS: In total, 135 853 patient and 62 080 donor samples were analysed for ABO typing discrepancies. The incidence among patients and blood donors was found to be 0·1% (138/135853) and 0·02% (14/62080), respectively. The mean age for patients and blood donors was 48·4 and 29·2 years, respectively. The most common cause of ABO typing discrepancies was due to cold autoantibodies among the patients (50·7%) and blood donors (57%) causing discrepant results in reverse typing. The various other causes of reverse typing discrepancies among patients were weak/missing antibody (25·4%), cold-reacting alloantibody (4·3%), warm autoantibody (2·2%), anti-A1 antibody (2·2%), Bombay phenotype (1·5%), transplantation (0·7%) and rouleaux (0·7%), whereas in blood donors, the causes were cold-reacting antibody (7%) and weak antibody (7%). The major cause of forward typing discrepancies among patients (12·3%) and blood donors (29%) was ABO subgroups. CONCLUSION: The resolution of ABO typing discrepancy is essential to minimise the chance of transfusion of ABO-incompatible blood.

Double Dose Plateletpheresis: A Savior to Shrinking Donor Pool and Platelet Inventory Management.

The double dose plateletpheresis (DDP) is considered to be a cost effective way of preparing platelets, owing to the low incidence of infectious complications and by also minimizing allogeneic donor exposure to the patients. We aimed at collecting DDP at our center and study its effect on donor hematological parameters, evaluate the product quality and the adverse donor reactions thereafter. Double Dose Platelet was collected from 160 eligible apheresis donors on Amicus cell separator (Fenwal, Inc. Three Corporate Drive Lake Zurich, IL, USA). The donor hematological parameters, product yield, adverse effects on the donors, collection efficiency (CE) and collection rate of the machine were noted. A total of 160 DDPs were collected. The total blood volume processed to achieve the yield of 6.0 × 1011 was 3673.5 ± 276.56 mL. The average yield achieved was 6.14 ± 0.26 × 1011. The average run time was 68.05 ± 6.25 min. Total ACD used was 408.33 ± 33.81 mL. We observed significant relation of pre-donation donor platelet count and platelet yield (p < 0.001). The CE was 78.09 ± 5.15%. There was a significant drop in the post DDP platelet count (p < 0.01) causing no adverse effect. Fourteen donors (8.75%) experienced mild citrate related adverse events. DDP does not lead to major adverse effects and post DDP hematological parameters are also within the acceptable range. It also helps to maintain apheresis platelet inventory, reduce donor exposure, reduce donor requirement and reduce the cost of the product.

Role of Anti-MICA Antibodies in Graft Survival of Renal Transplant Recipients of India.

INTRODUCTION: The MIC (MHC class I chain-related) genes are a group of nonclassical MHC genes, located in the MHC class 1 region of chromosome 6. The aim of the present study was to find the prevalence of MHC class 1 chain-related (MICA) alloantibodies in patients undergoing live-related donor renal transplantation and its role in short-term graft survival. The role of blood transfusion in the formation of these antibodies was also studied. MATERIALS AND METHODS: Pretransplant samples of patients undergoing renal allograft transplantation were tested for anti-MICA antibodies. Association of various demographics, HLA-A + B + DRB1 mismatches, anti-HLA antibody screen, and anti-MICA antibodies was assessed using Pearson's chi-square test. RESULTS: Out of 646 serum samples, 94 (14.6%) were positive and 552 (85.4%) were negative for anti-MICA antibodies. Patients with anti-MICA antibody had a graft survival 89.3% as compared to 94.7% in patients without anti-MICA antibody (P < 0.05). The hazard ratio for all patients was 3.0701 (P < 0.05). Out of the 340 patients with no HLA antibodies, the presence of anti-MICA antibodies without any HLA antibodies (n = 43) was associated with poor outcome in the patients (hazard ratio of 2.768, P < 0.05). The presence of MICA antibodies with HLA antibodies did not decrease the graft survival (hazards ratio of 1.3750, P > 0.05). CONCLUSION: Preformed MICA antibodies independently increase the risk of kidney rejection and therefore recommend that guidelines should be formed for mandatory testing of these antibodies prior to renal transplant.

The good, the bad, and the ugly of luminex donor-specific crossmatch.

The presence of donor-specific antibodies directed against human leukocyte antigen significantly influences renal transplant because of antibody-mediated rejection. We performed the screening of pre-renal transplant patients for preformed anti-HLA antibodies using anti-human globulin augmented-complement-dependent lymphocytotoxicity crossmatch (AHG-CDCXm), luminex donor-specific crossmatch (LumXm) and HLA antibody screening. Seven hundred and fifty-four patients were assessed for LumXm. HLA antibody screening was possible in 325 out of 754 cases. All the three investigations viz. CDCXm, HLA antibody screening and LumXm was performed in 325 patients. All CDCXm positive patients (10/325, 3.08%) were also positive with LumXm and HLA antibody screen whereas 14 cases (4.31%) with CDCXm negative were positive with luminex-based assays. LumXm and HLA antibody screening were both positive in 24 (7.38%) cases, LumXm and HLA antibody screening were both negative in 275 (84.63%) cases and LumXm negative and HLA antibody screening was positive in 22 (6.76%) cases. However, there were four cases (1.23%) which were positive in LumXm in spite of being negative in HLA antibody testing. Single Antigen Bead (SAB) assay was performed in all patients positive for HLA antibody test. We suggest that LumXm is a useful and sensitive technique for the detection of anti-HLA antibodies in pre-transplant renal patients. However, other measures such as luminex antibody screen, SAB assay, history of the donor, and the class of antibodies involved should be taken into consideration for pre-transplant work up of renal patients.

Facts and Fallacies of Kidd Antibodies: Experience in a Tertiary Care Hospital in North India.

We have analyzed the method used in our laboratory to detect the most elusive, clinically significant alloantibody: the Kidd alloantibodies and find the most convenient procedure. A retrospective analysis of the method used in our laboratory for determining Kidd alloantibodies from January 2013 to May 2015 was conducted. The details of the event that sensitized the patient for red cell antibody formation and procedure used to detect the alloantibody were retrieved from the departmental records. Of 405 red cell antibody identification cases, 24 (5.9 %) had Kidd antibody (anti-Jka in 12: 50 % cases; anti-Jkb in 4: 16.7 % cases; multiple antibodies in 8: 32 % cases). Thirteen of 24 patients (54.2 %) had autocontrol positive of which 6 cases needed adsorption procedures whereas antibody/ies could be identified without adsorption procedure in the remaining 7 cases. All the 7 cases had autocontrol of 1+ strength. Of the 11 patients (45.8 %) with autocontrol negative, the antibody was identified using solid phase in 7 cases whereas tube panels were also used in the remaining 4 cases. Kidd alloantibodies though deceptive can be identified by sensitive techniques like the solid phase and simple but laborious techniques using the tube cell panels. Depending upon the reaction strength of the autocontrol, the routine autoadsorption process may be skipped and tube cell enzyme treated cells or solid phase techniques be used to get the results.

Efficacy of single, extended and goal directed immunoadsorption in ABO incompatible living related donor liver transplantation.

BACKGROUND: Liver transplantation is one of the solid organs most commonly being transplanted across the world. The indications, affordability and accessibility have grown manifold. To increase the donor pool, inclusion of ABO incompatible liver donors is being considered. To enhance the graft functioning and survival, immunoadsorption apheresis to reduce the ABO hemagglutination titres are on the rise. CASE REPORT: We report three cases ABO incompatible liver transplantation with immunoadsorption protocol. The patients were in poor general condition with Child-Turcotte-Pugh (CTP) class 'C' and there were no suitable ABO compatible grafts at the time. For all three cases, immunosuppressive protocol consisted of induction with Rituximab, followed by tacrolimus, mycophenolate mofetil and corticosteroids. The third patient received basiliximab for induction, in addition to the above protocol. First 2 patients received 1 immunoadsorption (IA) session with Glycosorb ABO® system (Glycorex AB, Sweden), pre-operatively. The third patient received 2 IA sessions pre-operatively. Baseline IgM plus IgG titres were 1024; 64 and 1024 for anti-B, anti-A and anti-B for patient 1, 2 and 3 respectively. Target pre-operative antibody titre was ≤16. Average post-operative length of stay was 17.3 days. There were no acute rejection. None of them, required any post-operative plasma exchange. CONCLUSION: Immunoadsorption is effective in reducing hemagglutination titres in recipients of ABO incompatible donor liver.

Human leucocyte antigen Class I and II alleles associated with anti-hepatitis C virus-positive patients of North India.

PURPOSE: Humans are the only known natural hosts of hepatitis C virus (HCV). This study was undertaken to examine the frequencies of human leucocyte antigens (HLAs) Class I and Class II genotype profiles in anti-HCV-infected patients of Northern India. MATERIALS AND METHODS: From a period of January 2013 to August 2014, 148 anti-HCV-positive patients of North India referred to the Department of Molecular Biology and Transplant Immunology, Indraprastha Apollo Hospitals, New Delhi, for performing HLA typing were included in the study. RESULTS: AFNx0102, AFNx0131 allele frequency decreased significantly in anti-HCV-positive patients. Frequencies for HLA-B loci did not reach any statistical significance. Among the Class II alleles, HLA-DRB1FNx0103 and HLA-DRB1FNx0110 were significantly higher in the patient population, and HLA-DRB1FNx0115 was significantly decreased in the patient population as compared to the controls. CONCLUSION: HLA-AFNx0133 was significantly increased as compared to control population and showed geographic variation in HCV-infected individuals of India.

Seroprevalence of infectious markers & their trends in blood donors in a hospital based blood bank in north India.

BACKGROUND & OBJECTIVES: Hepatitis B virus (HBV), human immunodeficiency virus (HIV), hepatitis C virus (HCV) and syphilis infections pose a great threat to blood safety. This study was undertaken to investigate the seroprevalence of serologic markers for transfusion transmitted infections (TTIs) among blood donors at a hospital based blood centre in north India over a period of nine years. METHODS: The results of serologic markers for TTIs (HBsAg, anti-HCV, anti-HIV and syphilis) of all blood donations (both voluntary and replacement) at our hospital from January 2005 to December 2013 were screened. Additional analysis was conducted to examine the prevalence trends associated with each of the positive marker. RESULTS: The data of 180,477 donors [173,019 (95.86%) males and 7,458 (4.13%) females] were analyzed. Replacement donations [174,939 (96.93%)] represented the majority whereas, only 5,538 (3.06%) donations were from the voluntary donors. The risk of blood being reactive was three times higher in male donors when compared with the female donors. The risk of blood being reactive for one or more infectious markers was 2.1 times higher in replacement donors when compared with the voluntary donors. Seropositivity of HIV, HBsAg, HBcAb, syphilis showed a significant decreasing trend (P<0.05) while there was an increasing trend in HCV infection which was insignificant. INTERPRETATION & CONCLUSIONS: This study reflects that the risk of TTIs has been decreased over time with respect to HIV, HBV and syphilis, but the trends for HCV remains almost the same in blood donors. Blood transfusion remains a risk factor for the spread of blood-borne infections. Therefore, improvements are needed to strengthen both safety and availability of blood.

Detection of alloimmunization to ensure safer transfusion practice.

BACKGROUND: Serological safety is an integral part of overall safety for blood banks. Emphasis is on the use of routinue Red Blood Cell (RBC) antibody screen test, at set time intervals, to reduce risks related to alloantibodies. Also emphasis is on importance of issuing antigen negative blood to alloantibody positive patients. Effect of using leucodepleted blood on the rate of alloimmunization is highlighted. The concept of provision of phenotypically matched blood is suggested. MATERIALS AND METHODS: Antibody screen test is important to select appropriate blood for transfusion. Repeat antibody screen testing, except if time interval between the earlier and subsequent transfusion was less than 72 hours, followed by antibody identification, if required, was performed in patients being treated with repeat multiple blood transfusions. Between February 2008 and June 2009, repeat samples of 306 multi-transfused patients were analyzed. Search for irregular antibodies and reading of results was conducted using RBC panels (three-cell panel of Column Agglutination Technology (CAT) and two cell panel of the Solid Phase Red Cell Adherence Technology (SPRCAT). Specificities of antibodies were investigated using appropriate panels, 11 cell panel of CAT and 16 cell panel of SPRCA. These technologies, detecting agglutination in columns and reactions in solid phase, evaluate the attachment of irregular incomplete antibody to antigen in the first phase of immunological reaction more directly and hence improve the reading of agglutination. Three to four log leuco reduced red blood cells were transfused to patients in the study using blood collection bags with integral filters. RESULTS: Alloimmunization rate of 4.24% was detected from 306 multiply transfused patients tested and followed up. The Transfusion therapy may become significantly complicated. CONCLUSION: Red cell antibody screening and identification and subsequent issue of antigen negative blood have a significant role in improving blood safety. Centers that have incorporated antibody screen test and identification have ensured safe transfusion. Identified patients should be flagged in a database and information shared. Such patients can be given carry-on cards and educated about the names of the identified antibodies. Full red cell phenotyping of individuals, patients and donors, can be feasibility.

Preoperative predictors of blood component transfusion in living donor liver transplantation.

CONTEXT: Extensive bleeding associated with liver transplantation is a major challenge faced by transplant surgeons, worldwide. AIMS: To evaluate the blood component consumption and determine preoperative factors that predict the same in living donor liver transplantation (LDLT). SETTINGS AND DESIGN: This prospective study was performed for a 1 year period, from March 2010 to February 2011. MATERIALS AND METHODS: Intra- and postoperative utilization of blood components in 152 patients undergoing LDLT was evaluated and preoperative patient parameters like age, gender, height, weight, disease etiology, hemoglobin (Hb), hematocrit (Hct), platelet count (Plt), total leukocyte count (TLC), activated partial thromboplastin time (aPTT), international normalized ratio (INR), serum bilirubin (T. bilirubin), total proteins (T. proteins), albumin to globulin ratio (A/G ratio), serum creatinine (S. creatinine), blood urea (B. urea), and serum electrolytes were assessed to determine their predictive values. Univariate and stepwise discriminant analysis identified those factors, which could predict the consumption of each blood component. RESULTS: The average utilization of packed red cells (PRCs), cryoprecipitates (cryo), apheresis platelets, and fresh frozen plasma was 8.48 units, 2.19 units, 0.93 units, and 2,025 ml, respectively. Disease etiology and blood component consumption were significantly correlated. Separate prediction models which could predict consumption of each blood component in intra and postoperative phase of LDLT were derived from among the preoperative Hb, Hct, model for end-stage liver disease (MELD) score, body surface area (BSA), Plt, T. proteins, S. creatinine, B. urea, INR, and serum sodium and chloride. CONCLUSIONS: Preoperative variables can effectively predict the blood component requirements during liver transplantation, thereby allowing blood transfusion services in being better prepared for surgical procedure.

Seroprevalence of anti-HCV antibodies among blood donors of north India.

BACKGROUND & OBJECTIVES: Transfusion of blood and blood products although considered as a life saving treatment modality, but may lead to certain infectious and non-infectious complications in the recipients. The purpose of this analysis was to monitor the seroprevalence of anti-HCV antibody in the blood donor population in a hospital based blood bank in north India, to evaluate the trends over the years (2001-2011). METHODS: Relevant information of all the blood donors who donated whole blood at the department of Transfusion Medicine, Indraprastha Apollo Hospitals, New Delhi from the January 1, 2001 to December 31, 2011 was retrieved from the departmental records. The number of donors who were found reactive for anti-HCV anatibodies was calculated. RESULTS: Of the 2,06,022 blood donors, 1,93,661 were males and 12,361 were females. The percentage of whole blood donors found seroreactive for anti-HCV antibodies was 0.39 per cent (n=795). The seroprevalence of anti-HCV in male blood donors was 0.38 per cent (n=750) and the respective seroprevalence in female blood donors was 0.36 per cent (n=45). No significant change in the trend of HCV seroprevalence was observed over the period under consideration. Maximum seroprevalence of anti-HCV was observed in the age group of 18 to 30 yr (0.41%) and the minimum in the age group of 51 to 60 yr (0.26%). INTERPRETATION & CONCLUSION: HCV seroprevalence in our study was 0.39 per cent and a decreasing trend with age was observed. No significant change in the trend of anti-HCV seroprevalence was seen over a decade. Since, no vaccine is presently available for immunization against HCV infection, transfusion transmitted HCV infection remains a potential threat to the safety of the blood supply.

Rapid-prenatal diagnosis through fluorescence in situ hybridization for preventing aneuploidy related birth defects.

BACKGROUND AND OBJECTIVE: Women with high-risk pregnancies are offered prenatal diagnosis through amniocentesis for cytogenetic analysis of fetal cells. The aim of this study was to evaluate the effectiveness of the rapid fluorescence in situ hybridization (FISH) technique for detecting numerical aberrations of chromosomes 13, 21, 18, X and Y in high-risk pregnancies in an Indian scenario. MATERIALS AND METHODS: A total of 163 samples were received for a FISH and/or a full karyotype for prenatal diagnosis from high-risk pregnancies. In 116 samples both conventional culture techniques for getting karyotype through G-banding techniques were applied in conjunction to FISH test using the AneuVysion kit (Abbott Molecular, Inc.), following standard recommended protocol to compare the both the techniques in our setup. RESULTS: Out of 116 patients, we got 96 normal for the five major chromosome abnormality and seven patients were found to be abnormal (04 trisomy 21, 02 monosomy X, and 01 trisomy 13) and all the FISH results correlated with conventional cytogenetics. To summarize the results of total 163 patients for the major chromosomal abnormalities analyzed by both/or cytogenetics and FISH there were 140 (86%) normal, 9 (6%) cases were abnormal and another 4 (2.5%) cases were suspicious mosaic and 10 (6%) cases of culture failure. The diagnostic detection rate with FISH in 116 patients was 97.5%. There were no false-positive and false-negative autosomal or sex chromosomal results, within our established criteria for reporting FISH signals. CONCLUSION: Rapid FISH is a reliable and prompt method for detecting numerical chromosomal aberrations and has now been implemented as a routine diagnostic procedure for detection of fetal aneuploidy in India.

Prevalence of Rh, Duffy, Kell, Kidd & MNSs blood group antigens in the Indian blood donor population.

BACKGROUND & OBJECTIVES: Little data are available regarding the frequencies of the blood group antigens other than ABO and RhD in the Indian population. Knowledge of the antigen frequencies is important to assess risk of antibody formation and to guide the probability of finding antigen-negative donor blood, which is especially useful when blood is required for a patient who has multiple red cell alloantibodies. This study was carried out to determine the frequencies of the D, C, c, E, e, K, k, Fy(a), Fy(b), Jk(a), Jk(b), M, N, S and s antigens in over 3,000 blood donors. METHODS: Samples from randomly selected blood donors from Delhi and nearby areas (both voluntary and replacement) were collected for extended antigen typing during the period January 2009 to January 2010. Antigens were typed via automated testing on the Galileo instrument using commercial antisera. RESULTS: A total of 3073 blood samples from donors were phenotyped. The prevalence of these antigens was found to be as follows in %: D: 93.6, C: 87, c: 58, E: 20, e: 98, K: 3.5, k: 99.97, F(a) : 87.4, Fy(b) : 57.6, Jk(a) : 81.5, Jk(b) : 67.4, M: 88.7, N: 65.4, S: 54.8 and s: 88.7. INTERPRETATION & CONCLUSIONS: This study found the prevalence of the typed antigens among Indian blood donors to be statistically different to those in the Caucasian, Black and Chinese populations, but more similar to Caucasians than to the other racial groups.

Hepatitis B core antibody testing in Indian blood donors: A double-edged sword!

BACKGROUND: Until lately, anti-HBc antibodies were considered an effective marker for occult Hepatitis B virus (HBV) infection and have served their role in improving blood safety. But, with the development of advanced tests for HBV DNA detection, the role of anti-HBc in this regard stands uncertain. MATERIALS AND METHODS: Anti-HBc and HBsAg ELISA and ID-NAT tests were run in parallel on donor blood samples between April 1, 2006 and December 31, 2010 at the Department of Transfusion Medicine, Indraprastha Apollo Hospitals, New Delhi. A positive ID-NAT was followed by Discriminatory NAT assay. RESULTS: A total of 94 247 samples were tested with a total core positivity rate of 10.22%. We identified nearly 9.17% of donors who were reactive for anti-HBc and negative for HBsAg and HBV DNA. These are the donors who are potentially non-infectious and may be returned to the donor pool. CONCLUSION: Although anti HBc testing has a definite role in improving blood safety, centers that have incorporated NAT testing may not derive any additional benefit by performing anti-HBc testing, especially in resource-limited countries like ours.

Correlation between HER2 gene amplification and protein overexpression through fluorescence in situ hybridization and immunohistochemistry in breast carcinoma patients.

BACKGROUND: In India, the incidence of breast cancer has increased in the urban population, with 1 in every 22 women diagnosed with breast cancer. It is important to know the HER2/neu gene status for a better prognostication of these patients. AIM: The aim of this study was to compare the efficacy of fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) for determining HER2/neu alteration in breast carcinoma. MATERIALS AND METHODS: A total of 188 histologically proven breast carcinoma cases between the years 2007 and 2011 were retrospectively analyzed on the paraffin tissue sections by both IHC and FISH techniques. FISH for HER2/neu gene amplification was performed on cases where the IHC status was already known and the results were compared. RESULTS: A total of 64 (30%) patients were found to be amplified and the remaining 124 (65.9%) cases were found to be unamplified through FISH. Patients observed with 3+ reading on IHC were later confirmed as unamplified in 29.5% cases through FISH. CONCLUSION: It has been confirmed with the present study that IHC is a prudent first-step technique to screen tissue samples for HER2/neu gene status, but should be supplemented with the FISH technique especially in equivocal cases.

Evaluation of the red cell hemolysis in packed red cells during processing and storage.

INTRODUCTION: Storage of red cells causes a progressive increase in hemolysis. In spite of the use of additive solutions for storage and filters for leucoreduction, some amount of hemolysis is still inevitable. The extent of hemolysis, however, should not exceed the permissible threshold for hemolysis even on the 42(nd) day of storage. STUDY DESIGN AND METHODS: Eighty units of packed red cells, 40 stored in SAGM post leucoreduction and 40 in ADSOL without leucoreduction filters, were evaluated for plasma hemoglobin by HemoCue Plasma Hemoglobin analyzer on the day of collection and on the 7(th), 14(th), 21(st), 28(th), 35(th) and 42(nd) days thereafter. The hemoglobin and hematocrit were also noted for all these units by the Beckman and Coulter analyzer. Percentage hemolysis was then calculated. OBSERVATIONS: Hemolysis progressively increased with the storage period in all the stored red cell units (SAGM as well ADSOL). However, on day 42(nd) of storage, free hemoglobin in all the red cell units was within the permissible level (which is 0.8% according to the Council of Europe guidelines and 1% as per the US FDA guidelines). The mean percentage hemolysis was slightly higher in the SAGM-containing bags with an integral leucoreduction filter as compared to the bags containing ADSOL. However this difference was marginal and not statistically significant. CONCLUSION: Hemolysis of the red cells increases with storage. However, maximum hemolysis does not exceed the permissible limits at any time thereby indicating the effect of optimum processing and storage conditions on red cell hemolysis.

Octaploidy in idiopathic thrombocytopenic purpura.

We report a case of an elderly 68-year-old male who presented in our hospital with chief complaints of petechial rashes and ecchymosis over extremities and bleeding from the oral cavity since 3-4 days prior to hospitalization. He saw a physician before coming to our hospital and received one dose of IV methylprednisolone and oral wysolone. He had come to our hospital for further management. Bone marrow karyotyping was done and chromosomal analysis revealed two cell lines. Eighty percent of the cells analyzed revealed apparently normal male karyotype. However, 20% cells analyzed revealed a total of 184 chromosomes, suggesting octaploidy.