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 Table of Contents  
ORIGINAL ARTICLE
Year : 2020  |  Volume : 5  |  Issue : 1  |  Page : 58-62

Role of extended red cell phenotyping in management of patient with multiple antibodies and their utility in development of indigenous cell panels for antibody screening


Prathama Blood Centre, Ahmedabad, Gujarat, India

Date of Submission10-Feb-2020
Date of Decision29-Feb-2020
Date of Acceptance17-Mar-2020
Date of Web Publication17-Apr-2020

Correspondence Address:
Ripal J Shah
Prathama Blood Centre, Ahmedabad, Gujarat
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/GJTM.GJTM_8_20

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  Abstract 


Background and Objectives: The frequencies of clinically significant blood groups antigens (Rh, Duffy, Kell, Kidd, MNS, P and Lewis) should be known to BTS to manage patients with clinically significant antibodies. This study was conducted to help develop in-house cell panels to manage such patients. Methods and Materials: A total of 331 donors with O blood group with age group of 18 to 45 years, repeatedly donating blood were included in the study. They were screened for antigens of Rh, Kell, Kidd, Duffy, MNS, P and Lewis blood group system. Results: Among 331 donors, 299 (90.3%) donors were RhD Positive. e antigen was prevalent in 328 (99.1%) donors. Only 2 (3.5%) donors with E antigen were lacking D antigen also. All D negative donors (9.7% of total donors) were having strong expression of c and e antigen on their red cells. In Kell system, 100% donors were k antigen positive. All K positive donors are also positive for D antigen. In Kidd and Duffy system, Jka and Fya are more prevalent. In Lewis system, Lea-Leb+ (66.5%) is the commonest phenotype. In MNS system, M antigen was present in 87.61% of donors and s antigen in 83.38% of donors. Conclusions: Knowledge of red cell antigen phenotype frequencies in a population is helpful in terms of their ethnic distribution, in creating a donor data bank for preparation of indigenous cell panels, and providing antigen negative compatible blood to patients with multiple alloantibodies.

Keywords: Alloimmunization, antibody, blood group, red cell phenotype, voluntary donor, red cell panel


How to cite this article:
Shah RJ, Harimoorthy V, Shah RB, Barot TK, Kumar KM. Role of extended red cell phenotyping in management of patient with multiple antibodies and their utility in development of indigenous cell panels for antibody screening. Glob J Transfus Med 2020;5:58-62

How to cite this URL:
Shah RJ, Harimoorthy V, Shah RB, Barot TK, Kumar KM. Role of extended red cell phenotyping in management of patient with multiple antibodies and their utility in development of indigenous cell panels for antibody screening. Glob J Transfus Med [serial online] 2020 [cited 2020 Oct 25];5:58-62. Available from: https://www.gjtmonline.com/text.asp?2020/5/1/58/282751




  Introduction Top


Karl Landsteiner truly opened the doors of blood banking with his discovery of the first human blood group system. Few years later, two significant discoveries were made that would further increase the safety of blood transfusion and eventually define the most extensive blood group system known.[1]

The “blood group” refers to the entire blood group system comprising red blood cell (RBC) antigens whose specificity is controlled by a series of genes which can be allelic or linked very closely on the same chromosome. “Blood type” refers to a specific pattern of reaction to testing antisera within a given system. At present, 38 blood group systems representing over 300 antigens are listed by the International Society of Blood Transfusion.[2],[3]

The ABO and Rh blood groups are the most significant blood groups in transfusion practice. However, there are over 300 RBC antigens that are formally recognized internationally.[4] The other important blood groups are Kell, MNS, Duffy, and Kidd. These are significant in routine transfusion medicine; antibodies to these antigens are more commonly encountered.

The common antigens of the Rhesus and Kell blood group systems are also frequently tested in blood donors and patients because of their clinical relevance. Several studies,[2],[3],[4],[5] conducted in various parts of India, documented the antigen prevalence for the major blood group systems.[5],[6],[7],[8],[9]

The blood transfusion services mainly test for ABO and RhD antigens during cross-matching, particularly in resource-limited countries.[10] Normally, it is considered secure and time saving, but for the multi-transfused patients (thalassemic and anemic), the extended phenotyping for the determination of minor antigens is also necessary. The frequencies of clinically significant blood group antigens (Duffy, Kell, Kidd, and MNS) should be known to Blood Transfusion Service (BTS) to manage patients with clinically significant antibodies.[10]

Aim and objective

The present study was conducted to find the prevalence of different blood group antigens in the donor population of the city, to prepare donor data bank for the development of future indigenous cell panels, and provide antigen-negative compatible blood unit to patients with single/multiple alloantibodies.


  Materials and Methods Top


The study was conducted for 10 months from September 2017 to June 2018. The data were analyzed retrospectively. During the study period, 23,724 whole blood donors donated blood voluntarily. All the donors were screened according to the national guidelines for blood donation in India, which, generally, considered age, medical history, infections, and screening of donors.[11] All blood units were subjected to routine blood testing including transfusion-transmissible infection and blood grouping.

Sample inclusion criteria

Donors with O blood group were included in the study. The inclusion criteria used were donors with an age group of 18–45 years, repeatedly donating blood, ready to donate in future too as well as easily approachable (nearby places of the city). All donor samples included in the current study were selected only after confirming that their direct antiglobulin test results were negative.

Data collection

A total number of 331 donors were included in this study. Peripheral blood samples were collected in an ethylenediaminetetraacetic acid tube for ABO and Rh grouping as a routine test for blood donation. A barcode number was given to each donor at the time of blood donation. The samples were followed with donation ID barcodes. The ABO-Rh group was performed by fully automated grouping system (Neo, Immucor, Germany). The group O samples were further subjected to extended phenotype. The test was performed either using the conventional tube technique (181) or ID cards (150) and the results were then recorded in a datasheet. The samples were typed for D, C, c, K, k, Fya, Fyb, Jkb, Jka, Lea, Leb, P1, M, N, S, and s antigens using monoclonal antisera and E, e, antigens were typed using monoclonal blend antisera as per the manufacturer's instructions. For P1 and s antigens, antisera used were from Bio-rad, Switzerland, and the rest were from Immucor, US.

Statistical tool

Data were compiled in the Microsoft Excel format and analyzed accordingly.

Preparation of blood samples

For all ID-Cards except for ID-Card Antigen Profile III, a 5% red cell suspension was prepared and mixed gently, in a suspension tube using 0.5 ml of ID-Diluent 1 (Diamed Ag, Switzerland) and 25 μl of packed cells. The red cell suspension was incubated for 10 min at room temperature. After incubation, the cell suspension would be used within 15 min. As for ID-Card Antigen Profile III, a 0.8% red cell suspension was prepared in a clean tube using 1.0 ml of ID-Diluent 2 (modified LISS) (DiaMed AG, Switzerland) and 10 μl of packed cells and mixed gently. The cell suspension would be used immediately.

Test methods

The test was performed as per the manufacturer's instructions. The procedures are carried out for different sets of antigens. For IgG antigens (D, C, E, c, e, K, k, Fya, Fyb, Jka, Jkb, S, and s), the test was performed at AHG phase at 37°C, and for IgM antigens (M, N, P1, Lea, and Leb), the test was performed at saline phase. The antigens of Rh systems were mainly checked with standard tube technique using our standard operating procedures, while other IgG antigens were checked with column agglutination technology (CAT). The antigens reacting at the AHG phase were performed in CAT, while the rest were checked with standard tube technique.

Quality control

The quality of the antisera used in the study was checked using known antigen-positive and antigen-negative red cells present in commercial red cell panels.

Ethical clearance

The present study was retrospective. Donor details were made confidential. Local management's clearance was obtained before the data compilation.


  Results Top


During the study, a total of 331 random voluntary blood donors with group O were tested for Rh, Kell, Duffy, Kidd, Lewis, P, and MNS phenotypes.

Among the selected donors, 299 (90.3%) donors were RhD positive, while 32 (9.7%) were RhD negative [Figure 1]. Looking into antigen of minor Rh system, e antigen was widely prevalent in 328 (99.1%) donors, followed by D antigen and C antigen in 299 (90.3%) donors and 274 (84%) donors, respectively [Figure 2]. Only 57 (17.2%) donors had E antigen on their red cells. Only 2 (3.5%) donors with E antigen lacked D antigen, while the rest 55 (96.5%) donors also had D antigen.
Figure 1: Prevalence of RhD antigens

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Figure 2: Prevalence of minor Rh system antigens

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In the Kell blood group system, only 9 (2.7%) donors were typed as K antigen positive and 100% donors were k antigen positive [Figure 3].
Figure 3: Prevalence of Kell blood group system antigens

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In the Kidd blood group system, Jka antigen was prevalent in 232 (70.1%) donors and Jkb antigen was prevalent in 171 (51.7%) donors. Jka+Jkb− was prevalent in 37.8%, while Jka+Jkb+ was prevalent in 32% of donor population [Figure 4].
Figure 4: Prevalence of Kidd blood group system antigens

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In the Duffy blood group system, Fya was present in the majority of donors. Among 331 donors, 258 (84.5%) donors had Fya antigen on their red cells [Figure 5]. Looking into phenotypes of Duffy blood group system, 48 donors had Fya-Fyb+ phenotypes, while none of the donor was Fya−Fyb[Figure 6]. Fya+Fyb+ and Fya+Fyb− were present in 44.1% and 41.4% of the donors [Figure 6].
Figure 5: Prevalence of Duffy blood group system antigens

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Figure 6: Prevalence of Duffy blood group system phenotypes

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In the Lewis blood group system, the most prevalent phenotype was Lea−Leb+ (66.5%) followed by Lea+Leb−. Lea+Leb+ phenotype was found in very few donors. Only 2.4% of the donors had Lea+Leb+ phenotype [Figure 7].
Figure 7: Prevalence of Lewis blood group system phenotypes

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In MNS blood group system, M+N−S+s+ was the most common phenotype (28.4%) followed by M+N+S+s+ (17.52%) and M+N−S−s+ (16.01%) phenotype. Among our study group donors, none of the donor had S-s phenotype.

P1 antigen was present in 179 donors (54.1%).


  Discussion Top


The incidence of antigens of various blood groups in the local donor population helps in routine blood transfusion practices of a blood center.[12] The knowledge of prevalence of different blood group antigens in any given population is always helpful in managing cases of alloimmunization. Multiply transfused patients such as those with thalassemia and sickle cell anemia, patients on dialysis, and cancer patients are likely to develop antibodies against these minor blood group antigens as it is not practically feasible to match all these minor antigens before transfusion to avoid immunization. Finding compatible units for such patients without having any knowledge of the prevalence of the implicated antigens in the local population is a difficult task, more so if the patient has developed more than one antibody.

In India, various studies have been carried out to check for the prevalence of blood group phenotypes over a period of time. In different studies, D antigen was found from 84.35% to 96.6% of the population.[13],[14],[15],[16],[17],[18] In the present study, D antigen was found in 299 (90.3%) donors of 331 donors, which is comparable from other studies while higher than the study from the same region. The present study was conducted only in one center as compared to the cited study which was conducted in three different centers of the region. This could be the reason for variation in the prevalence of D antigen.

The frequency of C antigen in the present study was 84% which is comparable to 84.76% in a study in North Indian blood donors by Thakral et al. The frequency of E antigen was comparable in our donor populations and North Indian donors (17.2% and 17.6%, respectively). The frequency of e antigen in the present study was 99.1%, comparable to 100% found by Kahar andPatel and Shah et al. The strong association of C antigen was found with D antigen. Almost 99.3% of donor population where C antigen was present also had D antigen in the red cells, which is comparable to Kahar and Patel where 93.81% of donors had both the antigens. In the present study, all D-negative donors (9.7% of total donors) had strong expression of c and e antigen on their red cells, which is higher than Musa et al. where 7.5% of Indian donors had so. In Asians, approximately 3% of the Rh-negative populations had ce antigen.[19]

Different studies conducted about phenotypes have variable incidence rate for Kell system. K antigen was present from 1.6% to 6.09% of donors. In the Kell group system, k antigen was present in 100% of the donors which is comparable to Kahar and Patel[8] and Thakral et al. This may be different regions and different ethnic groups were studied in different studies. In the present study, K antigen was found in 2.7% of the donors, which was comparable to Yasmeen et al.[20] and Shah et al. (2.6% and 2.3%, respectively). In the present study, all K-positive donors also had D antigen.

In the Duffy system, Fya found in 85.5% of the donors and Fyb found in 58.6% of the donors. In the study by Makroo et al., the prevalences for both the antigens were 87.4% and 57.7% of the donors which was comparable to the present study. Fya-Fyb-phenotype was not present in any of the donor, which is comparable to Thakral et al.[6] In Kidd system, Jka+Jkb- was prevalent in 37.8%, while Jka+Jkb+ was prevalent in 32% of donor population, which is lower than found by Jabin et al.

In the Lewis system, Lea-Leb+ (66.5%) is the most common phenotype followed by Lea+ Leb-, comparable to that of Kahar and Patel and Thakral et al. In MNS system, M antigen was present in 87.61% of the donors, N in 46.22% of the donors, S antigen in 65.86% of the donors, and s antigen in 83.38% of the donors. The frequency of M and N antigen was comparable to Jabin et al, while S and s antigens were comparable to Makroo et al.

P1 antigen was present in 54.1% of the donors.

The prevalence of antigen varies among ethnic groups. The commercially available cell panel may miss the development of indigenous antibody developed, especially in multi-transfused patients, such as thalassemia major and sickle cell disease. Gujarat has high prevalence of thalassemia major patients.[21],[22] Repeated blood transfusion from multiple donors, thalassemia patients may develop of antibody/ies. Blood center having donors with known phenotype would always be useful in providing antigen-negative blood support to the patients with multiple alloantibodies or antibodies against antigen commonly found in the ethnic group.


  Conclusions Top


Knowledge about the prevalence of different blood group antigens in the donor population of the blood center, especially repeat voluntary blood donors, will be helpful to prepare donor data banks for the development of future indigenous cell panels. At the same time, it will be useful in providing antigen-negative compatible blood units to patients with single/multiple alloantibodies. The present study can be further extended for antigens of Indian origin which can be further utilized to antibodies to such antigens, which would further aid in the improvement of transfusion services in the country.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7]



 

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