Ankit Mathur

Serial follow-up of repeat voluntary blood donors reactive for anti-HCV ELISA

Background: Voluntary non-remunerated repeat blood donors are perceived to be safer than the first time blood donors. This study was planned for follow-up of previous hepatitis C virus (HCV) test results of anti-HCV enzyme-linked immunosorbent assay (ELISA) reactive repeat blood donors. The aim was to suggest a protocol for re-entry of the blood donors who are confirmed HCV negative by nucleic acid test (NAT) and recombinant immunoblot assay (RIBA). A group of repeat voluntary donors were followed retrospectively who became reactive on a cross sectional study and showed HCV reactivity while donating blood regularly. Material and Methods: A total of 51,023 voluntary non remunerated blood donors were screened for anti-HCV ELISA routinely. If anybody showed positivity, they were tested by two ELISA kits (screening and confirmatory) and then confirmed infection status by NAT and or RIBA. The previous HCV test results of repeat donors reactive by anti-HCV ELISA were looked back from the records. Data of donors who were repeat reactive with single ELISA kit (in the present study) were analyzed separately from those reactive with two ELISA kits (in the present study). Results: In this study, 140 (0.27%) donors who were reactive by anti HCV ELISA were included. Out of them, 35 were repeat voluntary donors and 16 (11.43%) were reactive with single ELISA kit. All 16 donors were reactive by single ELISA kit occasionally in previous donations. Their present ELISA positive donations were negative for HCV NAT and RIBA. A total of 19 (13.57%) donors were reactive with two ELISA kits. In their previous donations, the donors who were reactive even once with two ELISA kits were consistently reactive by the same two ELISA kits in their next donations also. Conclusion: Donor sample reactive by only single ELISA kit may not be considered as infectious for disposal as they were negative by NAT and or RIBA. One time ELISA positivity was found probably due to ELISA kit specificity and sensitivity. Donors reactive with two ELISA kit should be discarded as there is a high positivity with NAT/ RIBA. However, donors reactive by two ELISA kits and negative by NAT and RIBA should be followed up and may not be deferred permanently.

True positivity of anti-Hepatitis C Virus Enzyme-linked immunosorbent assay reactive blood donors: A prospective study done in western India

Background: A significant number of safe donations are removed from the blood supply, because of the reactive anti-HCV screening test results. This study aimed to assess if the HCV (Hepatitis C Virus) seropositive donors were confirmed positive or not. Materials and Methods: More than 68,000 blood donors’ samples were routinely screened and 140 samples were found to be anti-HCV ELISA reactive. These 140 samples were tested by NAT. The NAT negative samples were tested by RIBA. Analysis of samples reactive in single ELISA kit vs. two ELISA kits was done. Results: Out of 140 anti-HCV ELISA reactive samples, a total of 16 (11.43%) were positive by NAT. The results of 124 RIBA showed 6 (4.84%) positive, 92 (74.19%) negative, and 26 (20.97%) indeterminate results. None of the sample which was reactive in only single ELISA kit was positive by NAT or RIBA. Conclusion: Only a minority of blood donors with repeatedly reactive anti-HCV screening test is positive by confirmatory testing, but all these blood units are discarded as per existing legal provisions in India. Efforts should be made to retain these donors and also donor units.

Analysis of efforts to maintain safe donor in main donor pool after completion of temporary deferral period

Background: Voluntary blood donation is not satisfactory all over India. In India, about 55% of donation is through voluntary non-remunerated blood donors (VNRBD). However, about one third already motivated blood donors are deferred due to stringent screening criteria, either temporarily or permanently. The temporarily deferred donors could be a good source of blood donation after deferral period. Aims: The present study is carried out to know retrieval of blood donors those who are deferred temporarily. Design: The present study is carried out in the Regional Blood Transfusion Centre of Western India. All donors screened as per the guideline and deferred donors are categorized as temporary and permanently deferred donors. Materials and Methods: From temporarily deferred donors, reason for deferral is considered. As per reason of deferral, time duration for recalling the donor is defined. Based on this, donor is called back to donate again. Statistical Analysis: Chi-square test is applied. Result: A total of 33% donors were deferred either temporarily or permanently. In the repeat donors (5.32%) deferral rate was significantly higher than first time (1.32%) donors. Significant female preponderance was observed (15.05% vs 2.51%). Majority of temporarily deferred donors were less than 40 years of age (80.80%), graduate (82.90%), from low income group (62.90%) and profession was service (48.10%). Conclusion: Low hemoglobin (78.30%) was the most common reason of temporary deferral, both in first time and repeat donors (71.00%). Efforts to increase the hemoglobin in the repeat donors will improve the donor retention and overall blood safety can be increased.

Visual detection of hemolysis in a blood bag before issue

Hemolysis may occur in a red blood cell (RBC) unit during blood collection, transportation, preservation and or different stages of handling in the blood bank. Visual detection of hemolysis of a unit is possible usually by observing the color of the supernatant plasma. It is a good practice to observe the color of the plasma just before issue to avoid any inadvertent serious transfusion of a hemolysed blood unit. However, practically it is difficult once the stationary blood unit is disturbed or there is no time to centrifuge every unit of cross-matched blood to observe the plasma color.

Deferral pattern in voluntary blood donors on basis of low hemoglobin and effect of application of digital hemoglobinometer on this pattern

Background: One of the responsibilities of blood center is to provide safety to blood donors. It is mandatory to screen a blood donor for hemoglobin (Hb) or hematocrit which should not be less than 12.5 g/dl or 38% Hct. Most commonly applied method for hemoglobin estimation is copper sulphate method, but this method has chances of false acceptance as well as false deferral. In order to avoid this chance of error, digital hemoglobinometer is used. This study was planned to analyze effect of application of digital hemoglobinometer for detection of Hb on donors, who are deferred by copper sulphate method. Materials and Methods: Total 35,339 voluntary non renumareted altruistic donors were included in this study between the periods of September 2005 to July 2006. Total deferred donors were 8622 (24.39%) and donors deferred due to hemoglobin by copper sulphate method were 4391 (50.92%). Digital hemoglobinometer was applied on 3163 deferred donors (72.03%). Results of digital hemoglobinometer were validated by known controls. Result: Digital hemoglobinometer was applied on 3163 donors who were deferred by copper sulphate method. Out of this, donors accepted by digital hemoglobinometer were 1196 (37.01%). Total repeat donors were 629 (52.50%) and first time were 567 (47.40%). Male donors were 891 (74.44%) and females were 305 (25.50%). Donors deferred with digital hemoglobinometer were 2135, out of them 1097 (51.14%) were repeat, 1038 (48.38%) were first time, 1349 (60.79%) were male, 786 (34.47%) donors were female donors. Range of hemoglobin in deferred donors was 7.0 to 12.4 and in accepted donors 12.5 to 16.4. Conclusion: By the application of digital hemoglobinometer 37.81% donors were found hemoglobin >12.5 which were deferred with copper sulphate method and unnecessary deferral of donors can be reduced to a great extent. In country like India, where blood supply is always less than the requirement, this new technique may be helpful to increase donor population but cost benefit ratio should be analyzed.

Red Blood Cell Alloimmunization in Multi - transfused Patients: A Bicentric Study in India

Background: It is well-known that alloimmunization to red blood cell antigens resulting from the genetic disparities between donor and recipient is one of the risks of blood transfusion. The antibody screening cells are used to detect unexpected antibodies. The risk of alloimmunization is higher in patients who have received multiple blood transfusions such as thalassemia, other hematological disorders, renal failure patients on dialysis who receive blood transfusions, and females with bad obstetric history. Antibody screening test using 2–3 cells panel is not a mandatory pretransfusion testing in India and is performed routinely in limited blood centers. Materials and Methods: This bicentric study has been carried out to look at prevalence of antibodies in multi-transfused patients who have higher risks of alloimmunization such as patients who have received multiple blood transfusions such as thalassemia, other hematological disorders, renal failure patients on dialysis who receive blood transfusions, and females with bad obstetric history in the North as well as South India. The study was conducted at two blood centers, one regional blood transfusion of North India and one at South India. Totally, 4569 cases were analyzed and 258 patients were selected for antibody screening and identification. Results: Of total 4569 patients, 258 multi-transfused patients were studied. Among these, seven patients (2.71%) were found alloimmunized. The risk of alloimmunization was 2.90% in thalassemia, 0% in chronic renal failure patients, 3.77% in pregnant females with bad obstetric history, and 2.78% in other multi-transfused patients like cancer. Discussion: Regular monitoring through antibody screening and transfusion of blood matched for minor erythrocyte antigens are recommended in these patients.

An unusual case of a potentially clinically significant anti-M antibody in a healthy male blood donor without any history of blood transfusion

Dear Sir, The prevalence of red blood cell (RBC) alloantibodies among general, hospital-based patients typically has averaged approximately 1% in various studies. The presence of irregular antibodies in healthy voluntary blood donors is uncommon. Anti-M is a fairly common, naturally occurring antibody. Most anti-M are not active at 37 oC and generally are ignored in transfusion practice1. Sometimes, however, they can be clinically significant and become a problem in the immunohaematology laboratory. A male volunteer donor age 32 years donated blood in August 2008. Blood cell grouping showed an “A” positive phenotype, but serum grouping showed a positive reaction (+3) with all the three reagent red cells: A cells, B cells and O cells. The tests were repeated on the ID-Card, DiaClon ABO/D + reverse grouping (gel card) and the same results were observed. When reverse grouping tubes were tested at three different temperatures, 4 oC, 22 oC and 37 oC, for 1 hour, the reaction remained the same at 22 oC and 37 oC but negative at 4 oC. Autocontrols and the direct antiglobuling test were negative. With an antibody screening cell panel, two out of three cells showed positive results and an antibody identification panel (ID-DiaPanel 1-11 by DiaMed, Gmbh 1785 cressier, FR, Switzerland) showed that the suspected antibody was anti-M. The most likely type of antibody was IgG as the plasma was reactive even after treatment with dithiothreitol, which destroys IgM antibodies2. The A1 and O cells used for reverse grouping were shown to be positive for the M antigen. The anti-M present in the sample was interfering in reverse grouping with A1 and O cells. The A1 and O cells treated with papain enzyme gave a negative reaction in reverse grouping, as papain destroys the M antigen. This further confirmed the presence of anti-M with a wide thermal range, reacting at room temperature as well as at 37 oC. The antibody should, therefore, be considered potentially clinically significant. In blood donors, the prevalence of naturally occurring anti-M detectable in microplates with saline suspended cells at room temperature is one in 2,500 with M+N– cells and one in 5,000 with M+N+ cells 3. Some anti-M antibodies are pH-dependent and react at the optimum pH of 6.5. Another feature of this antibody is its failure to react with ficin- or papain-premodified cells. Proteolytic enzymes, such as ficin and papain, cleave the red cell membrane at well-defined sites. Haemolytic disease of newborn, of varying degrees of severity, has been reported in association with anti-M4,5.

A study of centralized individual donor nucleic acid testing for transfusion transmitted infections to improve blood safety in Karnataka, India

Introduction: Karnataka state has a total of 176 blood banks, with a total collection of around 650,000 units annually. From January 2014, all units under the Department of Health and Family Welfare services are tested at NAT Lab established at the Central facility and from September 2014, standalone Regional Blood Transfusion Centre was included in the State Government project. Aim and Objective: The aim of the study is to analyze the nucleic acid testing (NAT) for our donor population and demonstrate consolidation of blood transfusion service through a centralized testing center for NAT and also to assess safety benefits of implementing individual donor NAT (IDNAT). Materials and Methods: We collect nearly 40,000 units annually from voluntary donors with 30% repeat donations. The donors undergo strict predonation counseling, donor questionnaire, and medical examination. The units collected are tested for human immunodeficiency virus (HIV), hepatitis B virus (HBV) and hepatitis C virus (HCV) by ECI using VItros 3600. All the units are tested by NAT at Central NAT Lab, screened by Procleix® Panther System by Grifols. Results: From September 2014 to March 2016, total 50,903 samples tested for NAT. Of 50,903 samples, 588 samples (1.15%) were reactive by Chemiluminescence including 265 for HBV, 188 for HCV, and 135 for HIV. Total NAT reactive samples were 254, out of this 11 reactive for HIV-1 (0.02%), 2 reactive for HCV (0.003%), 235 (0.46%) reactive for HBV. There was one HIV and 10 HBV infection cases that were not detected by serology but reactive by NAT. The yield detected is 0.021% or one in 5000. Conclusion: The IDNAT project has helped in preventing window period infections thus reducing the treatment cost and burden on healthcare. It has added benefits in blood safety and should be considered along with the basic quality assured blood transfusion system such as volunteer base for blood donation, provision of donor self-deferral, donor notification, and quality assured sensitive serological methods.