Determination of optimum levels of binding antibody units (BAU) of new quantitative chemiluminescent immuno-assay (CLIA) in COVID-19 vaccinated volunteer blood donors.

BACKGROUND: For assessment of COVID-19 vaccine efficacy, neutralization activity of anti-SARS-CoV-2 antibody is measured. This study was undertaken to determine optimum levels of binding antibody units (BAU/ml) in new quantitative chemiluminescent assay (CLIA) that corresponded to neutralizing potential (30% inhibition) of sVNT assay. METHODS: Ninety-one blood samples were analyzed by CLIA and sVNT assays. Test samples (n = 75) were collected from blood donors post-2nd vaccination dose, while control samples (n = 16) were archived pre-COVID donor samples. Correlation between CLIA and sVNT was calculated and receiver operating characteristic (ROC) curve was drawn and analyzed. RESULTS: Results indicated excellent correlation between 57.5 BAU/ml on CLIA and 30%inhibition on sVNT assay. ROC curve analysis revealed that the area under the curve (AUC) was 0.971. DISCUSSION: The present study determined that 57.5 BAU/ml on CLIA corresponded to 30% inhibition on sVNT assay. Periodic quantitative analysis.

I-JAMM-(I): A survey providing an insight into the practices of isoagglutinin titration in ABO incompatible kidney and liver transplantation.

BACKGROUND AND OBJECTIVES: 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. METHODS: 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: 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. CONCLUSION: 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.

A patient with anti-f in India identified by extensive immunohematologic workup.

Anti-f is produced by exposure to the compound antigen ce (f) on red blood cells (RBCs), expressed when both c and e are present on the same protein (cis position). Although anti-f was discovered in 1953, there are few cases reported worldwide because the presence of anti-f is often masked by anti-c or anti-e and is not generally found as a single antibody. In the present case, anti-f was identified by using three-cell screening and 11-cell identification panels. The identification of anti-f was further supported by additional testing, including (1) Rh antigen typing; (2) antibody identification panels (enzyme-treated panel [ficin] and an in-house-constructed Rh panel); (3) look-back and phenotyping of donor RBC units, which were responsible for alloimmunization; and (4) molecular testing of the patient's RBCs.

Comparison of in-vitro and in-vivo parameters of whole blood derived random donor platelets (RDP) after over-night hold with RDP prepared after 2-h hold: Single centre report from India.

BACKGROUND: Random Donor Platelet (RDP) derived from whole blood is the major source of platelets in India. At our centre, we prepare RDPs by buffy coat method after a holding period of 2-hours (THRDP) as per current regulatory guidelines. Overnight hold of buffy coats before RDP preparation (OHRDP) would logistically optimise the manpower usage at our centre. The aim of this study was to compare both in-vitro as well as in-vivo parameters of OHRDPs with THRDPs. METHODOLOGY: Hematological (Platelet, leucocyte counts), physical (pH and Swirling) and biochemical parameters (pO2, pCO2, lactate, bicarbonate and glucose) as well as platelet activation markers were tested in THRDPs and OHRDPs each at Day-1 and Day-5 as in-vitro studies. Separately, in-vivo study was done where Corrected count increment (CCI) and percentage platelet recovery (PPR) were considered. All parameters were expressed as Mean ± Standard deviation and were analysed using paired t-test with level of significance, p < 0.05. RESULTS: OHRDPs had higher platelet counts and lower leucocytes and CD62 P expression than THRDPs. All other markers were well within the quality control range in both groups. No significant differences were seen in the two groups when comparing CCI and PPR. CONCLUSION: OHRDPs were found to be as good or better as compared to the THRDPs in the in-vitro part of our study. Additionally, there were no significant differences between the two groups when they were compared in vivo. This makes us conclude that overnight hold of buffy coats may be implemented at our center.

The frequent and the unusual red cell phenotypes in Indian blood donors: A quest for rare donors.

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.

Safety assessment of RhD-positive red cell transfusion in RhD-negative liver-transplant recipients: Single-centre report from India.

BACKGROUND & OBJECTIVES: The number of blood components required during a liver-transplant surgery is significant. It is challenging for blood transfusion services to provide the required RhD-negative red blood cells (RBCs) for recipients during the peri-operative period. This retrospective study presents safety data of transfusing RhD-positive RBCs in RhD-negative living donor liver-transplant (LDLT) recipients during the peri-operative period with six-month follow up for risk of developing alloantibodies. METHODS: All RhD-negative patients who underwent LDLT and were transfused ABO-compatible but RhD-positive RBC units between January 2012 and May 2018 were included in the study. Twenty one RhD-negative patients who received a total of 167 RhD-positive RBCs peri-operatively were chosen for alloantibody screening. All the patients were started on triple immunosuppression drugs as per the standard hospital protocol. Blood grouping, cross-match and antibody screening were done by column agglutination technique. RESULTS: Post-transplant antibody screen (weekly for 12 wk) was negative, and none of the patients developed anti-D alloantibodies till their last follow up (mean 21 months). INTERPRETATION & CONCLUSIONS: Our observations suggest that it may be safe to use RhD-positive RBCs peri-operatively in RhD-negative LDLT recipients with low risk of alloimmunization.

Automated-red cell exchange for methaemoglobinaemia in a G6PD-deficient patient.

Methaemoglobinaemia in G6PD deficiency can be managed by oxidizing agents such as methylene blue and red cell exchange (RCE). We describe a G6PD-deficient patient who presented with oxidative stress with methaemoglobinaemia and was successfully managed with automated-RCE. At presentation, the patient had anaemia, was restless, was tired and had dyspnoea. Co-oximetry showed methaemoglo-binaemia of 10.1 U/g. Further testing revealed the patient had insufficient quantities of G6PD enzyme activity (0.1 U/g Hb). In view of methaemoglobinaemia, severe G6PD deficiency and signs of haemolysis, therapeutic RCE was planned. The patient underwent two automated-RCE procedures on consecutive days, bringing down his methaemoglobin levels from 12.5 to 0.1 U/g. In each procedure, 1.5 volumes of RCE at 100% balance rate was performed using 5 units of red blood cells. The patient responded well to RCE and other supportive treatment and was off medication and doing well at day 100 of follow-up.

Applying newer parameter Ret-He (reticulocyte haemoglobin equivalent) to assess latent iron deficiency (LID) in blood donors-study at a tertiary care hospital in India.

BACKGROUND: It is important to detect Latent Iron Deficiency (LID) to prevent development of an overt iron deficiency anemia. Early detection is difficult by using conventional hematological and biochemical parameters. Soluble transferrin receptor (sTfR) is presently the gold standard for diagnosing LID. We evaluated the utility of Reticulocyte Hemoglobin Equivalent (Ret-He), a newer hematological parameter, to predict LID in blood donors as compared to sTfR. METHODS: This was a randomized prospective study performed on 501 donor samples over a period of three-months. All donors were included after administering medical history questionnaire and a brief physical examination in accordance with national guidelines (Hb ≥12.5). Additional samples were collected during donation according to the institutional standard operating procedure (SOP). All hemograms were performed on the Sysmex XE-2100 analyzer which included Ret-He. sTfR was measured in batch assays by ELISA (Biovendor, Czech Republic). Ret He <28 pg and sTfR≥3µg/ml were used to diagnose LID. Serum Iron, Total Iron Binding Capacity (TIBC) and Serum Ferritin were also measured simultaneously. RESULTS: Of the 501 blood donors, sTfR and Ret-He detected LID in 148 and 135 donors respectively. In comparison to sTfR, Ret-He had sensitivity of 92.7%, a specificity of 97.16%, PPV of 93.1% and NPV of 96.3%. Serum Ferritin, TIBC and serum Iron had comparatively lower sensitivity of 87.16%, 79.7% and 77.7% respectively. CONCLUSION: Ret-He can be used as a routine screening test to detect LID in blood donors. This could provide an opportunity to make appropriate and timely interventions like dietary changes or drug supplementation.

Notification and counselling of hepatitis positive blood donors, their immediate emotional response, contact-testing and their follow-up: Study from a tertiary care hospital!

INTRODUCTION: Post-donation counselling informs donors of unusual test results. Timely notification and counselling regarding their Transfusion Transmitted Infection (TTI) status is necessary for early clinical intervention in the donor and reducing risk of transmission. We share our experience with respect to Hepatitis B (HBV) and Hepatitis C (HCV) positive donors who were counselled and followed-up for clinical outcome. MATERIALS AND METHODS: It was prospective 2-year study in TTI positive blood donors. Confirmed positive HBV/ HCV donors were notified to attend the donor-clinic or to visit local hepatologist for further management. At donor clinic, donor's immediate emotional response was observed; donors were offered contact-testing, associated risk factors were noted, counselled, referred to hepatologist, treated and followed-up for clinical outcome. RESULTS: Of 481 donors (0.91%) confirmed positives, 351 were contacted telephonically; 280 promised to attend donor clinic and 71 were referred to their local hepatologist. 145 donors attended the donor clinic, eventually. Most common immediate emotional response noted were 'feeling of fear' (55.2%) and 'disbelief' (35.2%). Most common associated risk factor was history of medical treatment/ injections without knowledge of sterilisation. Five donors availed contact testing and four (spouses in all four cases) came out positive. Of 98 donors contacted post-counselling; 89 went to hepatologist. No medication was advised to seven donors (low viral load), 59 donors completed treatment course and 23 donors were undergoing treatment at time of follow-up. Nine donors opted for alternative treatment or "no treatment". CONCLUSION: Donor-clinic proved beneficial to substantial number of donors and their families.

Post-donation telephonic interview of blood donors providing an insight into delayed adverse reactions: First attempt in India.

BACKGROUND: Blood donor experiences both immediate adverse reactions (IAR) and delayed adverse reactions (DAR). With limited published data available on the incidence of DAR, a study was conducted to estimate incidence and profile of DAR through telephonic interview. MATERIALS AND METHODS: Study was conducted over a 45-day period for consecutive volunteer whole blood donations at tertiary care hospital. Donors were divided into first-time, repeat and regular and were monitored for IAR. They were given written copy of post-donation advice. Donors were contacted telephonically three weeks post-donation and enquired about general wellbeing and specific DAR in accordance with a standard n international (International Society of Blood Transfusion) standard format. RESULTS: Donors participated in the study of which 1.6% donors experienced an IAR. Much larger number reported DAR (10.3% vs.1.6% p<0.0001). Further, DAR was presented as a variegated profile with bruise, painful arms and fatigue being the commonest. DARs were more common in females than males (25% vs. 10.3%, p<0.02). Localized DAR like bruise and painful arms were more common in younger donors (age <50 years) whereas systemic DAR like fatigue was common in older donors (>50 years). First time (12.3%) and repeat donors (13.5%) had similar frequency of DAR but were lower among regular donors (6.7%). CONCLUSION: DARs are more common than IAR and are of different profile. Post-donation interview has provided an insight into donor experiences and can be used as a valuable tool in donor hemovigilance.

Co-infection of blood borne viruses in blood donors: A cross-sectional study from North India.

BACKGROUND: There are several studies on prevalence of individual infectious disease markers (mono-infection) in donors but none on prevalence of coinfection. Co-infection is significant as it leads to accelerated disease progression. We, therefore, evaluated the prevalence of co-infection among blood donors. MATERIALS AND METHODS: The cross-sectional analysis was conducted in blood donors. All donors were tested for anti-HIV I and II, HBsAg, anti-HBC IgM, anti-HCV, Malaria and syphilis by chemiluminescence and ID-NAT assay. All reactive donor samples were confirmed by using confirmatory assays. Donors were grouped as mono-infected and co-infected. The student t-test was used for comparison. RESULTS: During the study period, a total of 106,238 blood donors were tested. Mean age of donors was 34.2 years and 94.2% of blood donors were males. 1776 (1.67%) donor samples were confirmed serologically reactive. 1714 (1.61%) samples were reactive for single marker (mono-infected) while 62 (0.05%) donors' samples exhibited co-infection. 18 donors were positive for HBV+HCV followed by HIV +syphilis (14). CONCLUSION: We report for the first time the prevalence of different co-infection patterns in blood donors. Co-infection influence the disease progression; it would be important to investigate the co-infection prevalence in larger sample size.

Applications of selected cells in immunohematology in a developing country: case studies.

CONCLUSIONS: When an antibody is detected, its specificity should be determined and its likely clinical significance should be assessed. When one antibody has been identified, it becomes necessary to confirm the presence of additional significant antibodies to ensure that compatible blood is provided to the patient. To perform this confirmation, specific reagent red blood cells (RBCs) are selected; these are called selected cells. Though the most common use of selected cells is for antibody confirmation, they can also be used for several other immunohematologic applications. In a developing country like India, the performance of antibody screening for unexpected antibodies on a routine basis is a comparatively new phenomenon, and those laboratories performing advanced immunohematologic testing would need to use selected cells to arrive at an accurate conclusion. This report defines selected cells and enumerates sources of these RBCs. Detailed immunohematologic applications are discussed with applicable case studies.

Successful tandem transplant in a young aplastic anemia patient from a small-weight 11-month-old sibling donor.

Five years five month old male child diagnosed with aplastic anemia required blood and platelet support regularly. He was advised bone marrow transplant (BMT) and had 6/6 match with a younger sibling (11 months old). He was admitted for planned BMT and was put on preparatory regimen five days prior to BMT. GCSF-primed bone marrow (BM) harvest was done from the donor, but the harvest was insufficient (1.05 × 10^6 /kg) and target dose of 4 million stem cells per kg could not be achieved. The BM harvest was infused into the patient and a repeat BM harvest was contemplated on next day. After careful evaluation of risks and benefits to the patient and the young donor, a decision to do a peripheral blood stem cell (PBSC) harvest was taken. The apheresis kit was blood primed to preclude possible hemodynamic imbalance in the donor considering low weight and young age. The entire harvest procedure (321 minutes) was uneventful with the donor remaining stable throughout. A dose of 2.40 million per kg of CD34+ cells was harvested and infused. Thus, a total dose of 3.45 million (1.05 BM and 2.4 PBSC) per kg was infused into the patient. Neutrophil and platelet engraftments and donor chimerism were achieved successfully with tandem BM-PBSC infusion and the patient continues to be disease free till 180 days follow up. This is possibly first published Indian report on BM-PBSC tandem transplant suggesting safety of PBSC harvests in small-weight young-age donor.

Allo-antibody identification: a software approach!

BACKGROUND: Unexpected allo-antibody identification is difficult serological test requiring in-depth knowledge of antibody behavior, identification rules, knowledge of zygosity of antigens and dosage phenomenon. Software which uses an algorithm based on characteristics of antibodies is now available to interpret specificity of allo-antibody. A study was undertaken to evaluate the effectiveness of one such software (Resolvigen) for antibody identification compared with manual antibody identification method. MATERIALS AND METHODS: The study was a retrospective observational study where 238 allo-antibody results were re-evaluated using Resolvigen software (Ortho Clinical Diagnostics, Johnson and Johnson, Raritan, NJ, USA) and agreement between manual and software approaches was studied. Resolvigen software was also evaluated for usefulness, ease of use and predicted future usage by administering investigators a questionnaire with Likert scale. RESULTS: Agreement between the results of manual and automated methods ranged from 98.6% for single antibody to 65% for two antibodies (p = 0.000). Resolvigen software came out as very useful, easy to use, and with high predicted future usage. CONCLUSION: This study concludes that Resolvigen can either replace manual method or be used as adjuvant to routine manual method.

Anti-G with Concomitant Anti-D and Anti-C in a Pre-natal Patient, in India: Multi-disciplinary Approach to Diagnosis and Management.

'G' antigen belongs to the Rh family and it was first described by Allen and Tippet in 1958. Various anti-D, anti-C, and anti-G antibody combinations can be found in patients. Ruling out the presence of anti-D is important for administering RhIg prophylaxis in RhD-negative pregnant women to prevent hemolytic disease of fetus and newborn (HDFN). RhIg prophylaxis is not indicated in the presence of an anti-D antibody. Time-to-time monitoring and follow-up of cases of RhD-negative pregnant women with a multi-disciplinary approach including an obstetrician, neonatologist, and transfusion medicine specialist helps diagnose, manage, and monitor HDFN in such cases. This case report emphasizes the need for proper antibody identification (anti-G) and managing HDFN (with intrauterine transfusions and exchange transfusion) during the perinatal period.

Low-MFI (median fluorescence intensity) pre-transplant DSA (donor specific antibodies) leading to anamnestic antibody mediated rejection in live-related donor kidney transplantation.

"In solid organ transplantation, the compatibility between recipient and donor relies on testing prior to transplantation as a major determinant for the successful transplant outcomes. This compatibility testing depends on the detection of donor-specific antibodies (DSAs) present in the recipient. Indeed, sensitized transplant candidates are at higher risk of allograft rejection and graft loss compared to non-sensitized individuals. Most of the laboratories in India have adopted test algorithms for the appropriate risk stratification of transplants, namely: 1) donor cell-based flow-cytometric cross-match (FCXM) assay with patient's serum to detect DSAs; 2) HLA-coated beads to detect anti-HLA antibodies; and 3) complement-dependent cytotoxicity crossmatch (CDCXM) with donor cells to detect cytotoxic antibodies. In the risk stratification strategy, laboratories generally accept a DSA median fluorescence index (MFI) of 1000 MFI or lower MFI (low-MFI) as a negative value and clear the patient for the transplant. We present two cases of live-related donor kidney transplants (LDKTs) with low-MFI pre-transplant DSA values who experienced an early acute antibody-mediated rejection (ABMR) as a result of an anamnestic antibody response by DSA against HLA class II antibodies. These results were confirmed by retesting of both pre-transplant and post-transplant archived sera from patients and freshly obtained donor cells. Our examples indicate a possible ABMR in patients with low MFI pre-transplant DSA. Reclassification of low vs. high-risk may be appropriate for sensitized patients with low-MFI DSA."

Comparison of Two Different Serological Viral Marker Testing Assays for Screening of Apheresis Donors: Which Assay Provides Optimum Safety for Transfusion?

While whole blood testing has evolved over the years, viral marker testing for plateletpheresis donors is still performed by Rapid Diagnostic Tests (RDT). Aim of this study was to compare diagnostic accuracy of RDT and Chemiluminescence Immunoassay (CLIA) in serological testing for HBsAg, anti-HCV and anti-HIV antibodies. A prospective, analytical study was conducted in the department of Transfusion Medicine at a tertiary healthcare center in India between September 2016 and August 2018. Samples were simultaneously tested by CLIA, RDT and a confirmatory test. Sensitivity, specificity, negative and positive predictive values and mean time taken to report results were calculated. A total of 102 (1.48%) of the 6883 samples were found to be reactive by either or both the assays. A total of 74 (1.08%) samples were HBsAg reactive, 23 (0.33%) were reactive for anti-HCV antibodies and 5 (0.07%) were reactive for anti-HIV I and II antibodies. A combined sero-prevalence of 1.05% (72) was observed; 0.78% (54) for HBsAg, 0.26% (18) for anti-HCV antibodies and none for anti-HIV I and II antibodies. Four (3.85%) reactive samples were missed by RDT and therefore sensitivity of RDT was quite less as compared to CLIA. RDT and CLIA both were found to have a statistically significant shorter turnaround time than confirmatory tests. There is increasing need to develop a safe donor screening strategy for plateletpheresis. CLIA offers an excellent alterative to RDT for viral marker testing in terms of sensitivity.

Implementation of an international barcode labeling standard, International Society of Blood Transfusion 128, and its integration with local regulations at a blood center in India: Step-by-step journey.

The International Society of Blood Transfusion (ISBT) 128 is an internationally endorsed, electronically readable labeling standard that provides a convenient and accurate means of identification, traceability, publication, and storage of information for blood and blood products. The authors' center recently registered with the International Council for Commonality in Blood Banking Automation (ICCBBA) and progressed to ISBT 128 labeling standard. This manuscript was written with the objective of sharing the authors' experience with respect to the implementation of ISBT 128 standards for whole blood donations and integration of ISBT 128 standards with Indian licensing regulations. The authors explore the process of implementation of ISBT 128 standards through a step-by-step journey that included facility registration with International Council for Commonality in Blood Banking Automation (ICCBBA), allotment of facility identification number, development of four-quadrant label for blood components, and integration of local regulatory requirements in the final "composite" label. Acknowledging the lack of any published report from India on ISBT 128 standards implementation, the authors wish to attempt help their peers in understanding and implementation of this global standard at their respective facilities.

Analytical and clinical performance evaluation of enhanced chemiluminescence-based fourth-generation HIV combo assay: Report from tertiary health-care setup in North India.

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.

Real-world data on renal transplantations from a tertiary-care hospital in North India, in context of Indian regulatory act-transplantation of human organs and tissues act (THOTA): A retrospective analysis.

INTRODUCTION: Renal transplantation is the treatment of choice for patients suffering from end stage renal disease (ESRD). Indian regulations defined under Transplantation of Human Organs and Tissues Act (THOTA), 2014 restricts organ donations to near-related living donors to curb any malpractices like 'paid donors' in living-donor kidney transplantation (LDKT). The aim of our study was to look at real-world data of donor-recipient pairs and to identify relationship of donors (with their respective patients) and the common (or uncommon) DNA profiling methods used for supporting "claimed relationship" in accordance with the regulations. MATERIAL AND METHODS: The donors were categorized and grouped into near-related donor, donors other than near-related donors, swap donors and deceased donors. Claimed relationship was confirmed, commonly by HLA typing, using SSOP method. In few cases, which were uncommon (and infrequent), autosomal DNA analysis, mitochondrial DNA analysis and Y-STR DNA analysis were performed to support the claimed relationship. Data collected included age, gender, relationship, DNA profiling test method. RESULTS: Among the 514 donor-recipient pairs evaluated, numbers of female donors out-numbered male donors. The decreasing order of relationships in near-related donor group were wife>mother>father>sister>son>brother>husband> daughter>grandmother. 11.9% of donors were in the category of donors other than near-related donors. In 97.86% cases, the claimed relationship was supported by HLA typing and in just 2.1% cases autosomal DNA analysis>mitochondrial DNA analysis> Y-STR DNA analysis, in this order, were performed to establish relationship. CONCLUSION: This study brought out gender disparity with women out-numbering men as donors. Among recipients, access to renal transplant was largely restricted to men. As far as relationship of donors to recipients was concerned, mostly near-related family members, like wife, were donors and claimed relationship was almost always (99%) was corroborated by HLA typing.