Search

BACKGROUND AND OBJECTIVES: Enumeration of hematopoietic progenitor cell (HPC) is vital to decide the time to initiate harvest (TTIH) and adequacy of harvest dose (AOHD). Standard of care used for HPC enumeration is flowcytometric CD34+ enumeration, but it is expensive, time-consuming and requires skilled staff to perform the test. Alternatively, HPC-count by advanced automated cell analyzer is cheaper, quicker, and easy-to-perform test. Our objective was to find a correlation of HPC count with CD34+ enumeration in leukapheresis. MATERIALS AND METHODS: An observational, prospective study was conducted in the year 2018-2019. A total of 126 samples were included in the study, the peripheral blood (PB) group comprised of 42samples and apheresis group of 84 samples. The samples were simultaneously tested for CD34+ expression and complete blood count which included the HPC count, white blood cells (WBC) count and multinational corporation (MNC) count and correlation analysis was performed with CD34+ flowcytometric count. The cut-off of PB HPC count for the target dose of 5 x 106 CD34+ cells/kg was established using Receiver Operator Curve. RESULTS: The correlation coefficient (r) of HPC with CD34+ count was 0.617 and 0.699 for PB group and apheresis group sample respectively, which was statistically significant. The correlation with MNC and WBC count was not very significant. A cut-off value of PB HPC was established to be 66 HPC/µl with a positive predictive value of 94.12%. The cost of CD34 + flow cytometric enumeration was six times that of HPC enumeration by analyzer. CONCLUSION: The HPC count is a cheaper, rapid and easy test and can be clinically applied to predict TTIH and AOHD but requires more studies to validate its efficacy in clinical use. [ABSTRACT FROM AUTHOR]

INTRODUCTION: HIV fourth-generation assay, designed for the detection of HIV p24 antigen along with anti-HIV antibodies of both immunoglobulin M and immunoglobulin G type against HIV 1 and HIV 2 viral antigens, have helped in the early detection of HIV infection and supports in minimizing the transmission risk in the acute phase of infection. The objective of this study was to evaluate the analytical and clinical performance of HIV fourth-generation assay based on enhanced chemiluminescence technology. MATERIALS AND METHODS: The analytical performance of the assay was evaluated in terms of accuracy, precision, limit of detection, type of sample (serum vs. plasma), cross-reactivity (with other transfusion transmissible infections markers), and interference (with endogenous substances). Proficiency control material included kit-controls, archived known positive donor samples, third-party controls, and World Health Organization (WHO)/National Institute for Biological Standards and Controls (NIBSC, MHRA, UK) controls. The clinical performance was evaluated using routine donor and patient samples received during the study period. RESULTS: HIV fourth-generation assay showed reliable and reproducible results measured in terms of coefficient of variation % with kit-controls, archived known positive donor samples, third-party controls, and WHO international standards for anti-HIV 1 and 2 antibodies, HIV1 p24 antigens and HIV2 p26 antigen controls. The analytical sensitivity of the HIV fourth-generation assay was found to be 0.1 IU/mL of HIV1 p24 antigen control and there was no cross-reactivity or interference observed. In the clinical performance of the assay, HIV fourth-generation assay showed reliable performance in both donor and patient samples. CONCLUSION: HIV fourth-generation assay meets the requirements for its use as a screening assay for HIV infection based on the analytical and clinical performance of the assay. [ABSTRACT FROM AUTHOR]

BACKGROUND: In clinical practice, laboratory results are of great importance for the diagnosis and treatment. Reference intervals of different parameters aid health-care professionals in the interpretation of results. There are very few studies on reference intervals from India. This prospective study was conducted to determine the reference intervals for platelet count (PLT) and PLT indices; mean PLT volume (MPV), PLT distribution width (PDW), and PLT large cell ratio (P-LCR). These values can be obtained as a part of a routine complete blood count (CBC) and have diagnostic and prognostic significance in certain diseases. PLT count is an important criterion for the selection of donors for repeat plateletpheresis donation. MATERIALS AND METHODS: Sixteen hundred and thirty-four first-time healthy volunteer plateletpheresis donors were enrolled for the study. CBC was done, values of PLT, MPV, PDW, and P-LCR were noted, and the results were analyzed. The 95% of the reference distribution was estimated using the 2.5th and 97.5th percentiles following Clinical and Laboratory Standards Institute guidelines. Adverse donor reactions, if any and quality parameters of single donor PLTs (SDP) were also studied. RESULTS: Reference range values of PLT, MPV, PDW, and P-LCR were 137,825–355,175/μl, 8.1–13.9/fl, 9.1–22.5/fl, and 11.7%–52.9%, respectively, and compared well with other published studies from India. It was observed that reference values of PLT count obtained in the study were lower than reference values that are currently used in most laboratories (150,000–450,000/μl) in India. CONCLUSION: Based on our results, we are of the opinion that the PLT count cutoffs for repeat plateletpheresis donation may need to be revised downwards for our country which would also mitigate the scarcity of apheresis donors. [ABSTRACT FROM AUTHOR]

BACKGROUND: Human leukocyte antigen (HLA) is a major determinant in deciding upon solid organ histocompatibility. Donor-specific anti-HLA antibodies (Donor-specific anti-HLA antibodies (DSAs)) are always a contraindication for solid organ transplantation, and identification of DSA becomes very crucial before transplantation to provide long-term graft survival. For identification of DSA, usually, either cell-based or HLA bead-based assay is being used in laboratories. However, both cell-based and bead-based assays have certain limitations. One such common limitation is "prozone effect," which can give false-negative results. Here, we would like to present a small pilot study to analyze the effect of the prozone phenomenon in the cell-based and HLA bead-based assays and its utility in histocompatibility testing. MATERIALS AND METHODS: In a series of four experiments, cell-based assay, flow cytometric cross-match (FCXM), and HLA bead-based flow cytometric panel reactive antibodies (PRAs) were performed. Single-antigen bead (SAB) testing was conducted as a first experiment on four known positives samples for anti-HLA antibody-antibodies. In the second experiment, these four samples were pooled together (called pooled sera in the text) and tested for FCXM and PRA. In the third experiment, known commercially available positive control sera were mixed with pooled positive sera (positive control sera + pooled sera) to prepare, what we have called "positive concoction" in the text. In the fourth experiment, the positive concoction was diluted serially (1:2, 1:4, 1:8, and 1:16) and FCXM and PRA were performed again to analyze and compare the prozone effect. RESULTS: Pooled sera did not have the expected median fluorescence intensity (MFI) values in FCXM assay, whereas the PRA was showing >90% positivity. In positive concoction, the MFI of FCXM assay was observed to be declining; however, PRA values remained almost constant. Dilutions of the pooled sera showed that MFI values of FCXM assays were increased suddenly after dilution. The highest MFI values were observed in 1:4 dilution of the sera, and then, it declined gradually, but the PRA values remained almost constant even after serial dilutions. CONCLUSION: In our experimental findings, it was clear that cell-based assay (FCXM) was more severely affected by the prozone, whereas solid-phase (flow PRA) assay remained resistant to prozone. [ABSTRACT FROM AUTHOR]

OBJECTIVES: The study objective was evaluation of amotosalen and ultraviolet A (UVA) illumination-based inactivation of dengue virus (DENV) in blood platelets. MATERIALS AND METHODS: Whole blood was collected from healthy donors and platelet concentrates were prepared at a tertiary care hospital in Gurugram, India. Platelet units collected from five blood group matched individuals were pooled and spiked with DENV. The spiked platelet units were subjected to amotosalen treatment followed by UVA illumination, to evaluate the efficiency of this method for viral inactivation. The treated platelet units were evaluated for the presence of infectious DENV. Amotosalen levels were quantified in the treated samples using high-performance liquid chromatography. RESULTS: The presence of replicating DENV was not observed in spiked platelet units treated with amotosalen and UVA illumination, whereas untreated units contained actively replicating DENV. Amotosalen levels were found to be in the permissible range after photochemical inactivation. CONCLUSIONS: Amotosalen/UVA pathogen inactivation treatment showed efficient inactivation of DENV in platelet components. Therefore, it seems to be a promising method for mitigating the risk of dengue transmission through transfusion of potentially contaminated platelet components in dengue-endemic countries such as India. [ABSTRACT FROM AUTHOR]

Introduction: Solid organ transplantation is the preferred therapeutic modality of treatment in patients affected by terminal organ failures. Human leukocyte antigens (HLAs) plays an important role in graft survival. In many of the cases of rejection, antibodies are directed against HLA antigens expressed on the cells of the transplanted organ. Pre-transplant compatibility testing involves the use of different methodologies for the determination of anti-HLA antibodies. Luminex single-antigen bead (SAB) assay demonstrates higher sensitivity and specificity in detecting anti-HLA antibodies. The aim of this study was to determine the prevalence of anti-HLA antibodies in pre-transplant work up recipients, planned for renal transplant at a tertiary care center in India. Methods: 1640 patients visiting tertiary care hospital for pre-transplant compatibility testing were screened with complement-dependent micro-lymphocytotoxicity crossmatch (CDC-XM) and flow cytometric crossmatch (FC-XM). The patients positive for either or both screening tests were assayed with the Luminex SAB tests in order to establish defined antigen specificity of the alloantibodies and determining donor-specific antibody (DSA). Results: The two most frequent antibodies identified in each A, B, C locus of HLA class I were -A*24:03 (43.9%), A*25:01 (36.6%), B*57:01 (40.3%), B*15:12 (37.1%), C*17:01 (61.9%), C*07:01 (52.4%) and in DR, DQ DP locus in HLA class II were DRB1*09:01(40.0%), DRB1*14:04(37.6%), DQA1*04:01/DQB1*03:03 (58.4%), DQA1*05:01/DQB1*03:01 (55.1%), DPA1*02:01/DPB1*17:01 (55.0%), DPA1*02:01/DPB1*05:01 (45.0%). Conclusion: This study has found the prevalence and specificity of anti-HLA antibodies in north India. [ABSTRACT FROM AUTHOR]

Background: As a part of continuous quality initiatives, while moving from triple bags to quadruple bags, we undertook a study to compare platelet-rich plasma (PRP) and buffy-coat (BC) methods with respect to all blood components (red blood cells [RBCs], random donor platelet concentrate [RDPC], and fresh frozen plasma [FFP]) prepared by PRP and BC methods. Materials and Methods: It was a prospective analysis of different physical and quality parameters of RDPC, RBC, and FFP prepared out of 100 whole blood (WB) donations. Of these, 50 WB units were processed by PRP method using Triple bags and 50 WB units by BC method, using quadruple (top and bottom) bags, with an attached integral filter. Results: RBC prepared by BC method had higher hematocrit (61.3 ± 1.91% vs. 56.03 ± 3.37%; P < 0.05) and lower white blood cell (WBC) contamination (6.3 × 104 ± 6.1 vs. 5.41 × 105 ± 2.5; P < 0.05) in comparison to prepared by PRP method. Higher PLT yield (7.67 × 1010 ± 1.8 vs. 6.47 × 1010 ± 1.5; P < 0.05) and lower WBC count (8.24 × 103 ± 1.1 vs. 1.5 × 104 ± 2.1; P < 0.05) was observed in RDPC prepared by BC method than PRP derived. CD62P expression was lower in RDPC prepared by BC method (31.46 ± 9.7%; P < 0.05) as compared to PRP method (43.35 ± 12.5%; P < 0.05). The BC method also resulted in increased plasma yield (210.56 ± 18.54 ml vs. 187.92 ± 12.93 ml; P < 0.05) in FFP in comparison to PRP method. Conclusion: The blood components produced from WB by the BC method have laboratory variables suggestive of superior quality than those produced by the PRP method. [ABSTRACT FROM AUTHOR]

Background: Administration of anticoagulant citrate and dextrose (ACD-A) chelates ionized calcium in blood and causes hypocalcemia in plateletpheresis donors. The aim of the study was to observe the effects of oral calcium (Ca) supplementation during plateletpheresis on various parameters related to calcium metabolism. Materials and Methods: This study was performed between January 2014 and December 2014 on 200 plateletpheresis donors.They were divided into two groups. In group A donors (n=100), no prophylactic oral calcium supplementation was given. In group B (n=100) donors, 2000 mg of calcium was given one hour before the start of the procedure, 500 mg was given at the start of the procedure and 500 mg calcium was given just before the end of procedure. Biochemical parameters like serum total calcium (T Ca), serum total magnesium (T Mg) and ionized calcium level (iCa) were measured before and after the procedure. Relative risk of citrate toxicity was measured between the two groups. Results: There was a significant fall in total calcium (pre 9.02 mg/dl, post 8.23 mg/dl,), ionized calcium level (pre 1.14 mmol/L, post 0.91 mmol/L) and total magnesium (pre 1.92 mg/dl, post 1.79 mg/dl) amongst the donors who did not receive prophylactic calcium supplementation. Despite calcium intake, in prophylactic calcium intake group, we did observe a significant drop in total magnesium (pre 2.04 mg/dl, post 1.94 mg/dl) and ionized calcium level (pre 1.25 mmol/L, post 1.12 mmol/L, p<0.01). We did observe a drop in total calcium level, however, this observation was not statistically significant. The risk (RR=5.44) of citrate toxicity was higher among group A donors. Conclusion: Prophylactic oral calcium carbonate supplementation would help in to reduce the risk of citrate toxicity. Therefore, we suggest for prophylactic oral administration of 3000 mg elemental calcium carbonate in three divided doses to make PP procedures uneventful. [ABSTRACT FROM AUTHOR]

Background: Hematopoietic progenitor cell transplantation (HPCT) is used as a definitive treatment in hematological malignancies. For a successful HPCT, the donor and recipient should have matching human leukocyte antigens (HLAs). About 25% of patients have a chance of finding matching HLA within family, while rests 75% are dependent on voluntary stem cell donor. Globally, there are 75 stem cell registries with more than 30 million donors registered among which India represents 0.36 million. Therefore, finding a stem cell donor for Indian patient is quite difficult. The aim of the present study is to discuss the significance of voluntary stem cell donor recruitment drive and also to guide the drive organizers and their team for effectively organizing the drive to increase the database of such donors. Materials and Methods: Voluntary stem cell donor recruitment drives are conducted to spread awareness among the people and motivate them to register as a donor. Once the donors have given their consent, the sample is taken and sent to laboratory for HLA typing and the result is uploaded in World Marrow Donor Association, an international association of member to find the best possible matches for patients with hematological disorders. Results: Genebandhu has organized over 127 recruitment camps since 2012 and recruited 13,000 voluntary stem cell donors. HLA typing of 7446 donors has been completed. Out of this small number of typed donors, 11 lifesaving HPCTs have been successfully facilitated. Conclusions: Here, we have demonstrated guidelines along with steps to organize voluntary stem cell donors recruitment drive that is needed to increase number of donors, thus increasing significantly the chances of saving many vital lives. [ABSTRACT FROM AUTHOR]

Background and Objectives: The study was undertaken with the objective to provide data on the ABO and Rh(D) blood group distribution and gene frequency across India. Materials and Methods: A total of 10,000 healthy blood donors donating in blood banks situated in five different geographical regions of the country (North, South, East and Center) were included in the study. ABO and Rh (D) grouping was performed on all these samples. Data on the frequency of ABO and Rh(D) blood groups was reported in simple numbers and percentages. Results: The study showed that O was the most common blood group (37.12%) in the country closely followed by B at 32.26%, followed by A at 22.88% while AB was the least prevalent group at 7.74%. 94.61% of the donor population was Rh positive and the rest were Rh negative. Regional variations were observed in the distribution. Using the maximum likelihood method, the frequencies of the IA, IB and IOalleles were calculated and tested according to the Hardy Weinberg law of Equilibrium. The calculated gene frequencies are 0.1653 for IA (p), 0.2254 for IB (q) and 0.6093 for IO (r). In Indian Population, O (r) records the highest value followed by B (q) and A (p); O > B > A. Conclusion: The study provides information about the relative distribution of various alleles in the Indian population both on a pan-India basis as well as region-wise. This vital information may be helpful in planning for future health challenges, particularly planning with regards to blood transfusion services. [ABSTRACT FROM AUTHOR]

Background: In spite of many improvements that have reduced the blood component requirements, substantial numbers of transfusions are still needed in liver transplantation. Aims: The objective of the present study was to analyze the perioperative usage of allogenic blood components and predict the preoperative factors as predictors of red cell transfusion in live-related donor liver transplant recipients. Materials and Methods: The retrospective data on utilization of allogenic blood components were analyzed for a total of 150 liver transplant procedures. The data on utilization of blood components during surgery and till 48 hours of ICU stay was collected from the blood bank record and hospital information system (HIS). Results: Red cell concentrate was commonest blood component used in liver transplant recipient and most of the transfusion took place during surgery. During intraoperative period 92.7% (N = 139) of the cases utilized red cell components with the median number of five whereas in postoperative period only 38% (N = 57) of patients received blood with the median number of one. This study demonstrates that the preoperative hemoglobin and platelet count are the predictors of utilization of red cell concentrates during surgery. There were a total of 11 (7.3%) recipients who didn't receive allogeneic blood transfusion in any form.Utilization of blood components was negligible among organ donors. Conclusion: Our study demonstrates the pattern and predictors of usage of allogeneic blood components in liver transplant recipients at a tertiary healthcare center in India. [ABSTRACT FROM AUTHOR]