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 Table of Contents  
Year : 2019  |  Volume : 4  |  Issue : 2  |  Page : 219-223

Prevalence of irregular red blood cell antibodies among healthy blood donors in South India

1 Department of Transfusion Medicine, Dr. Rela Institute and Medical Centre; Department of Transfusion Medicine, Gleneagles Global Health City, Chennai, Tamil Nadu, India
2 Department of Transfusion Medicine, Gleneagles Global Health City, Chennai, Tamil Nadu, India
3 Department of Transfusion Medicine, Gleneagles Global Health City, Chennai, Tamil Nadu; Department of Transfusion Medicine, Apollo Gleneagles Hospital, Kolkata, West Bengal, India
4 Department of Transfusion Medicine, Dr. Rela Institutec and Medical Centre, Chennai, Tamil Nadu, India

Date of Submission06-Mar-2019
Date of Acceptance02-Apr-2019
Date of Web Publication17-Oct-2019

Correspondence Address:
Dr. G Deepthi Krishna
Department of Transfusion Medicine, Gleneagles Global Health City, Chennai, Tamil Nadu
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/GJTM.GJTM_22_19

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Background: Although antibody screening is mandatory according to the National Blood Policy of India, the prevalence of irregular red cell antibody and its specificities are not much reported in South Indian population. Aims and Objectives: The objective of this study is to find the prevalence of irregular red cell antibodies and positive direct antiglobulin test (DAT) in healthy blood donors in South India population. Materials and Methods: This is a retrospective observational study conducted in the department of transfusion medicine from April 2011 to March 2018 (7 years) at a tertiary care referral center, Chennai, Tamil Nadu. Red cell antibody screening of all voluntary blood donors was performed using commercial O cells (ID DiaCell Pool, DiaMed, Cressier sur Morat, Switzerland). Positive sera were further investigated to identify the irregular erythrocyte antibody by commercially available red cell panel (ID-DiaPanel, Diamed-ID Microtyping System). Results: A total of 40,629 donors were screened for the presence of irregular erythrocyte antibodies. A total of 57 (0.14%) donors showed the presence of irregular antibodies, in which 41 (71.9%) were alloantibodies. Most frequent alloantibodies identified were of MNS blood group system with anti-M being the highest (n = 15, 36.58%), followed by Rh blood group system. Sixteen (0.039%) donors were DAT positive, which were detected during routine crossmatching. Conclusion: Implementation of red cell antibody screening in all the blood donors routinely helped us understand the prevalence of antibodies in our region and its importance in providing compatible blood products and to avoid transfusion reactions.

Keywords: Antibody screening, blood donor, direct antiglobulin test, irregular antibody

How to cite this article:
Sachan D, Krishna G D, Saha S, Prasath R. Prevalence of irregular red blood cell antibodies among healthy blood donors in South India. Glob J Transfus Med 2019;4:219-23

How to cite this URL:
Sachan D, Krishna G D, Saha S, Prasath R. Prevalence of irregular red blood cell antibodies among healthy blood donors in South India. Glob J Transfus Med [serial online] 2019 [cited 2022 Jul 1];4:219-23. Available from: https://www.gjtmonline.com/text.asp?2019/4/2/219/269381

  Introduction Top

Irregular allo/autoantibodies refer to antibodies that are reactive at 37°C or in the anti-human globulin phase of testing. The reactivity of these antibodies is highly varied and unpredictable. These antibodies can be encountered in healthy blood donors who are either transfused previously or in multiparous females. As per the National Blood Policy, India, 2007, in addition to infectious marker screening, blood donors should be screened for irregular red cell antibodies to avoid any adverse transfusion reactions, especially due to the transfusion of plasma components.[1],[2]

There is a paucity of literature on the prevalence of the irregular red cell antibodies in whole blood donor population. The reported incidence of irregular antibodies from various studies ranges from 5 to 10 in 10,000 blood donors.[3],[4],[5]

Aims and objectives

To find the prevalence of irregular red cell antibodies and positive Direct antiglobulin Test (DAT) in healthy blood donors in South India population.

  Materials and Methods Top

This retrospective observational study was conducted inthe department of transfusion medicine of a tertiary care referral center, Chennai, Tamil Nadu, in accordance with the institutional ethical standards. Donors who were eligible according to the National AIDS Control Organization (NACO)guidelines were selected after donor consent and tested for the presence of irregular red blood cell (RBC) antibodies.

Meticulous donor screening was done at the time of blood donation which included donor basic profile, i.e., age, gender, address, any previous history of transfusion, any usage of drugs, history of jaundice or any significant disease associated to autoimmune disorders, and obstetric history in particular from female donors. Consent was taken before blood donation.

Universal precautions were taken during sample handling, processing, and testing. All the donor samples were collected from the donors during blood donation in ethylenediaminetetraacetic acid vials for blood grouping and antibody screening. Blood grouping was performed on semi-automated immunohematology system Saxo ID-centrifuge and ID-reader using commercially available DiaClon ABO/D+ Reverse Grouping (monoclonal antibodies) cards, Bio-Rad Laboratories. Antibody screening was performed using commercial single pooled “O” cells (ID-Diacell Pool, Bio-Rad Laboratories) with LISS/Coombs cards.

Any sample with positive antibody screen result using pooled “O” cells was subjected to antibody screening and identification with commercial screening reagent red cells (DiaMed, Switzerland) using commercially available cell panels (ID-DiaPanel, Diamed-ID Microtyping System). In addition to antibody screening, direct Coombs test was performed on donor samples in case of crossmatch incompatibility or blood group discrepancy, for example, reactions with O cells, as shown in [Table 1]. Thermal amplitude test was done in case of cold antibody to determine the nature of the antibody according to our standard operating procedure.
Table 1: The stage and method of initial donor antibody detection and antibody classification

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All the data was compiled, tabulated and frequencies were calculated using Microsoft Excel spread sheet.


All the data was compiled, tabulated and frequencies and percentages were calculated using Microsoft excel spread sheet.

  Results Top

A total number of 40,629 donors donated at our center during the study period, in which 96% of donors were male and 4% were female. A total of 57 donors showed the presence of irregular antibodies, in which 41 (71.9%) were alloantibodies and 16 were direct antiglobulin test (DAT) positive. [Table 2] shows the year-wise distribution of donations and prevalence of antibody positivity, in which there is an increasing trend of the prevalence of antibodies over the past 5 years. The prevalence of antibodies is higher in female donor population (0.5%) compared to males (0.12%), as shown in [Table 3]. Overall frequency of alloantibodies among blood donors is depicted in [Table 4]. Antibodies of MNS blood group system have the highest prevalence (51.2%) with anti-M (n = 15/41, 36.58%) being the most common antibody identified among the alloantibody donors, followed by anti-N (n = 5/41, 12.19%). Next common is the Rh system with anti-D (n = 4/41) showing 9.75%. When age-wise distribution of donors positive with antibody screen was considered, it shows that more number of donors (n = 31) belong to the age group of 25–44 years [Figure 1].
Table 2: Year-wise distribution of total number of donations and prevalence of antibody

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Table 3: Profile of total donors and antibody screen-positive donors

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Table 4: Frequency of alloantibody among antibody screen donors

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Figure 1: Age-wise distribution of donors with positive antibody screen

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  Discussion Top

Naturally occurring anti-A and anti-B antibodies are routinely detected in human serum and are easily detected by the use of A and B reagent RBCs with a direct agglutination technique. In normal healthy individuals, anti-A and anti-B are generally the only RBC antibodies expected to be found in serum. All other antibodies directed against red cells are unexpected and are acquired through transfusion or pregnancy. They are known to occur in general population as well. Factors influencing the immunization rate and distribution of RBC alloantibodies include age, gender, ethnicity, number of transfusions, autoimmune disease, immunoglobulin level, lymphoproliferative disease, solid malignancy, and amount of incompatible erythrocytes received by the individual through transfusion.[6]

Proper detection and identification of unexpected or irregular alloantibodies in donors are important for smooth and safer transfusion to patients. The prevalence of such irregular red cell antibodies has been extensively studied in patients in various parts of the world and also in India, but the data regarding exclusive donor population are very limited.

Data from various studies reported that the rate of alloimmunization in blood donors varies from 0.05% to 3.9%.[3],[4],[5],[6],[7],[8],[9],[10],[11],[12] In our study population, the prevalence of irregular antibodies is 0.14%, which is almost similar to other studies from India, as depicted in [Table 5]. The difference in various frequencies in various studies could be due to the different methodology used for antibody screening and identification and heterogeneity of the population studied. In our study, semi-automated column agglutination technology was used and it is less sensitive (94%) than the solid-phase red cell adherence assay method with a sensitivity of 97%, used by Makroo et al., which showed the highest prevalence among the studies.[10]
Table 5: Review of literature for the prevalence of red cell antibodies in donor population

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The study from Seattle, Washington,[13] had reported 0.32% incidence of irregular RBC antibodies in blood donors, and a study by Winters et al.,[6] in 2001, reported a prevalence of 1.4% (n = 257 of 18,185 donors studied) among blood donors of Olmsted County, Minnesota. This high prevalence of antibodies in these studies was attributed to high number of females in study donor population who were previously transfused or had a history of pregnancy.

In the present study, the highest frequency of alloantibodies was identified in blood donors aged between 25 and 44 years in our study, as shown in [Figure 1]. similar to other studies from North India,[5],[10] the high prevalence in this age group is due to maximum donors being in the reproductive age group and group comprising more number of active donors from this age.

In our study, the prevalence of irregular RBC alloantibodies in males (n = 48, 84.2%) was higher than females (n = 9, 15.7%) similar to a study by Makroo et al. (n = 63, 82.90%), correlating with the high prevalence of male donor population.[10] Naturally occurring antibodies which may present in either male or female population without a previous history of transfusion or pregnancy, if detected during antibody screening, should be checked for clinical significance as per institutional policies.

In the present study, 9.75% (n = 4) of donor antibodies could not be determined. Donors showing positive antibody screening with the presence of inconclusive antibody panel could be due to low-titer antibodies in the donor serum. The other possibility could be that these antibodies are directed to antigen not mentioned in the panel antigen profile.

High-prevalence antigens/high-frequency antigens (HFA) are those that are present in almost all individuals (98% or more).[1] They are suspected when screen and panel cells are positive, with reactions in the same phase and at the same strength, along with a negative autocontrol. Four donors in the present study (2.87%) showed the presence of anti-HFA antibodies in the serum. Those units are discarded as part of the departmental protocol, and donor notification was given. Need for reference laboratories is required to sort out the specificity of HFA antibodies. In case of patients, finding the compatible blood may be a challenge.

In our study, the most frequent alloantibodies identified were from the MNS blood group system, followed by Rh blood group system and Lewis system. The frequency of anti-M and anti-N was found to be 36.58% and 12.19%, respectively, which was clinically significant. Anti-M and anti-N are generally naturally occurring alloantibodies which do not react at 37°C and are not clinically significant for transfusion but can cause a problem in pretransfusion testing. These antibodies are clinically significant when reactive at 37°C where plasma and platelets should be discarded to prevent unusual transfusion reactions and packed red cells (PRCs) should be issued after appropriate crossmatch.[14],[15] The frequency of M antigen globally is 75%; hence, 25% of the individuals lack M antigen, who are able to generate anti-M antibody when exposed to the antigen which could be the best possible explanation of such high frequency.[16]

Next most common antibodies found are those of Rh and Lewis blood group systems. The frequency of antibodies of Rh blood group system in our study was found to be 17.07% (n = 7). Of these, anti-D was found to be 9.75% (n = 4), and the frequency of minor antigens anti-E and anti-D+C was found to be 7.3%.

The Lewis blood group system is different from other blood group systems, as the antigens (Lea and Leb) are formed in the plasma and absorbed onto the red cell membrane. Although Lewis antibodies present in recipient are implicated in Hemolytic Transfusion reaction, there is no report highlighting its clinical significance of Lewis antibodies in donor plasma[17] These antibodies may be clinically relevant if the antibody causes in vitro hemolysis during serologic laboratory testing and antigen-negative blood should be selected for transfusion[18] In our donors, anti-Leb (n = 4, 9.75%) was found to be more common than anti-Lea (n = 2, 4.87%). Like the present study, Chenna et al[12] also reported the presence of Lewis antibodies in their study group.

DAT may be positive in a small percentage of otherwise healthy donors.[19] A positive DAT in an otherwise healthy person might be considered to herald future autoimmune disease. Hence, it is equally important to test for DAT on all blood donors along with a screening of irregular antibodies. This adds an extra layer of safety in blood transfusion. In our study, 16 donors (n = 16/57, 0.039%) had autoantibodies (DAT positive) which are almost similar to results reported by Tiwari et al. (0.04%) and Kaur et al. (0.05%).[3],[11] As per our institutional policy, DAT-positive PRC units were discarded. There is also a need to do an elaborate long-term follow-up on these donors since there is an evidence of a significantly increased risk of cancer, especially hematologic malignancies, among blood donors with a positive DAT.[20]

The limitation of the study was incomplete immunohematology workups in cases of anti-HFA antibodies or inconclusive antibodies due to lack of availability of rare red cell antisera or local reference laboratories for serological or molecular confirmation. Irregular antibody screening in donors is not done in many centers in India due to decentralized and fragmented blood transfusion services. It serves to simplify the work of crossmatching by eliminating the need for the minor crossmatch. Red cell antibody screening of the donors is a simple test, adds a layer of safety in transfusion, and reduces the need for minor crossmatching for plasma transfusion. In such cases, only packed RBCs should be used for transfusion. In addition, we recommend that in cases where the antibody is found in blood donor, they should be informed, so that in future, if they require any transfusion, they can inform the blood bank prior.

  Conclusion Top

Data of the prevalence of red cell alloantibodies in blood donors will assist in improving safe blood supply for patients needing transfusion and to avoid transfusion reactions. We found that there is a significant frequency of irregular RBC alloantibodies (0.14%) with MNS blood group system being the most frequently identified alloantibodies in blood donors at our center. This kind of studies emphasizes the need for the mandatory screening and identification of irregular RBC antibodies among blood donors and it creates an eye view of the necessary for better use of donor blood, more efficient services as well as safe and compatible blood transfusion, especially for previously alloimmunized individuals.


D.S. conceived the study, designed and performed the research, participated in data collection, analyzed the data, and wrote the paper. R.P participated in data collection. D.K.G analyzed the data and wrote the paper, and S.S. participated in data collection, analyzed the data, and reviewed the paper.

We wish to express our gratitude to staff of blood bank for the technical support in performing the study.

Financial support and sponsorship


Conflicts of interest

There are no conflicts of interest.

  References Top

Harmening DM. Modern Blood Banking and Transfusion Practices. 6th ed. United States of America: FA Davis Company Publications; 2012.  Back to cited text no. 1
National AIDS Control Organization. National Blood Policy. India: Ministry of Health & Family Welfare; 2007. Available from: http://www.naco.gov.in/upload/Final%20Publications/Blood%20Safety/National%20Blood%20Policy.pdf. [Last accessed on 2019 Apr 12].  Back to cited text no. 2
Kaur D, Bains L, Kandwal M, Parmar I. Erythrocyte alloimmunization and autoimmunization among blood donors and recipients visiting a tertiary care hospital. J Clin Diagn Res 2017;11:EC12-5.  Back to cited text no. 3
Garg N, Sharma T, Singh B. Prevalence of irregular red blood cell antibodies among healthy blood donors in Delhi population. Transfus Apher Sci 2014;50:415-7.  Back to cited text no. 4
Pahuja S, Kushwaha S, Sethi N, Pujani M, Jain M. Screening of blood donors for erythrocyte alloantibodies. Hematology 2012;17:302-5.  Back to cited text no. 5
Winters JL, Pineda AA, Gorden LD, Bryant SC, Melton LJ 3rd, Vamvakas EC, et al. RBC alloantibody specificity and antigen potency in Olmsted County, Minnesota. Transfusion 2001;41:1413-20.  Back to cited text no. 6
Keokhamphoui C, Urwaijitaroon Y, Kongphaly D, Thammavong T. Red cell alloantibodies in Lao blood donors. Res Notes 2014;45:194-7.  Back to cited text no. 7
Sallander S, Shanwell A, Aqvist M. Evaluation of a solid-phase test for erythrocyte antibody screening of pregnant women, patients and blood donors. Vox Sang 1996;71:221-5.  Back to cited text no. 8
Ameen R, Al-Eyaadi O, Al-Shemmari S, Chowdhury R, Al-Bashir A. Frequency of red blood cell alloantibody in Kuwaiti population. Med Princ Pract 2005;14:230-4.  Back to cited text no. 9
Makroo RN, Rajput S, Agarwal S, Chowdhry M, Prakash B, Karna P. Prevalence of irregular red cell antibody in healthy blood donors attending a tertiary care hospital in North India. Asian J Transfus Sci 2018;12:17-20.  Back to cited text no. 10
[PUBMED]  [Full text]  
Tiwari AK, Pandey P, Sharma J, Shailja K, Dixit S, Raina V, et al. Incidence of clinically significant antibodies in patients and healthy blood donors: A prospective cross-sectional study from a tertiary healthcare center in India. Transfus Apher Sci 2014;50:230-4.  Back to cited text no. 11
Chenna D, Shastry S, Murugesan M. Significance of adopting a sensitive technique for donor antibody screening. Indian J Hematol Blood Transfus 2016;32:307-8.  Back to cited text no. 12
Giblett ER. Blood group alloantibodies: An assessment of some laboratory practices. Transfusion 1977;17:299-308.  Back to cited text no. 13
Fung MK, Grossman BJ, Hillyer CD, Westhoff CM. Technical Manual of American association of Blood Banks. 18th ed. Bethesda, Maryland: ABO, H, and Lewis Blood Groups and Structurally Related Antigens, Rh System and Other Blood Group System; 2014. p. 304-51.  Back to cited text no. 14
National AIDS Control Organization. Standards for Blood Banks & Blood Transfusion Services. India Ministry of Health & Family Welfare; 2007. Available from: http://www.naco.gov.in/upload/Final%20Publications/Blood%20Safety/Standards%20for%20Blood%20Banks%20and%20Blood%20Transfusion%20Services.pdf. [Last accessed on 2019 Apr 12].  Back to cited text no. 15
García MA, Bautista L, Palomino F. Should blood donors be routinely screened for irregular antibodies? Immunohematology 2012;28:60-6.  Back to cited text no. 16
De Vries SI, Smitskamp HS. Haemolytic transfusion reaction due to an anti-Lewis agglutinin. Br Med J 1951;1:280-1.  Back to cited text no. 17
Webert Kathryn E, Smith James W, Arnold Donald M, Chan Howard HW, Heddle Nancy M, Kelton John G. Red cell, platelet and white cell antigens. Wintrobe's Clinical Haematology. 12th ed. Lippincott Williams & Wilkins; 2009.  Back to cited text no. 18
Win N, Islam SI, Peterkin MA, Walker ID. Positive direct antiglobulin test due to antiphospholipid antibodies in normal healthy blood donors. Vox Sang 1997;72:182-4.  Back to cited text no. 19
Rottenberg Y, Yahalom V, Shinar E, Barchana M, Adler B, Paltiel O, et al. Blood donors with positive direct antiglobulin tests are at increased risk for cancer. Transfusion 2009;49:838-42.  Back to cited text no. 20


  [Figure 1]

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5]


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