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      A comparison of serological phenotyping and molecular genotyping for Kell, Kidd, and Duffy antigens in multi-transfused thalassemia patients

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          Abstract

          BACKGROUND:

          In multi-transfused thalassemia patients, serological phenotyping fails to test patient's actual blood group antigen profile due to the presence of donor red blood cell (RBC) in the circulation. This limitation of serological tests can be overcome by genotype determination using the polymerase chain reaction (PCR)-based methods. The aim of this study is to compare the serological phenotyping of Kell, Kidd, and Duffy blood group systems with molecular genotyping in the normal blood donors and multi-transfused thalassaemia patients.

          MATERIALS AND METHODS:

          Blood samples from 100 normal blood donors and 50 thalassemia patients were tested using standard serological techniques and PCR-based methods for Kell (K/k), Kidd (Jk a/Jk b), and Duffy (Fy a/Fy b) blood group systems. The results were compared for concordance.

          RESULTS:

          Genotyping and phenotyping results were 100% concordant for normal blood donors whereas those for thalassemia patients showed 24% discordance. The frequency of alloimmunization in thalassemia patients was 8%. The results of genotyping were used to provide Kell, Kidd, and Duffy matched blood for transfusion therapy to thalassemia patients.

          CONCLUSION:

          The actual antigen profile in multitransfused thalassaemia patients can be reliably determined using genotyping. This would benefit in providing better antigen matched transfusion therapy to such patients hence reducing the rate of alloimmunization.

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          Most cited references17

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          Molecular genetics and clinical applications for RH.

          Rhesus is the clinically most important protein-based blood group system. It represents the largest number of antigens and the most complex genetics of the 30 known blood group systems. The RHD and RHCE genes are strongly homologous. Some genetic complexity is explained by their close chromosomal proximity and unusual orientation, with their tail ends facing each other. The antigens are expressed by the RhD and the RhCE proteins. Rhesus exemplifies the correlation of genotype and phenotype, facilitating the understanding of general genetic mechanisms. For clinical purposes, genetic diagnostics of Rhesus antigens will improve the cost-effective development of transfusion medicine.
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            Alloimmunization and red cell autoimmunization in multitransfused thalassemics of Indian origin.

            Transfusion therapy in thalassemia is a double edged sword, on the one hand, it is a major life saving modality, while on the other hand, it can lead to complications such as alloimmunization. The rates of alloimmunization have been variably reported across the world; however, there is a paucity of such literature among Indian thalassemics. We studied the frequency of alloimmunization and autoimmunization among 211 multitransfused thalassemics of Indian origin. All the patients have been receiving blood matched for ABO and Rh(D) antigens only. Direct antiglobulin test was performed on all patients to detect autoantibody while antibody screening (using 3-cell panel) and antibody identification (11-cell panel) were carried out to detect the presence of alloantibody. The frequency of alloimmunization was 3.79% and that of autoimmunization was 0.47%. The alloantibodies identified were anti-E, anti-K, anti-D, anti-Kp(a), anti-C(w), anti-c and anti-Jk(a). In the present study, no significant association was observed between splenectomy and the development of alloantibodies as well as between age at initiation of transfusion and alloimmunization. To conclude, there is a need to formulate a balanced and cost-effective approach for transfusion management of thalassemics to minimize alloimmunization and autoimmunization.
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              Red cell genotyping and the future of pretransfusion testing.

              Over the past 20 years the molecular bases of almost all the major blood group antigens have been determined. This research has enabled development of DNA-based methods for determining blood group genotype. The most notable application of these DNA-based methods has been for determining fetal blood group in pregnancies when the fetus is at risk for hemolytic disease of the fetus and newborn. The replacement of all conventional serologic methods for pretransfusion testing by molecular methods is not straightforward. For the majority of transfusion recipients matching beyond ABO and D type is unnecessary, and the minority of untransfused patients at risk of alloimmunization who would benefit from more extensively blood group-matched blood cannot be identified reliably. Even if a method to identify persons most likely to make alloantibodies were available, this would not of itself guarantee the provision of extensively phenotype-matched blood for these patients because this is determined by the size and racial composition of blood donations available for transfusion. However, routine use of DNA-based extended phenotyping to provide optimally matched donations for patients with preexisting antibodies or patients with a known predisposition to alloimmunization, such as those with sickle cell disease, is widely used.
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                Author and article information

                Journal
                Asian J Transfus Sci
                Asian J Transfus Sci
                AJTS
                Asian Journal of Transfusion Science
                Wolters Kluwer - Medknow (India )
                0973-6247
                1998-3565
                Jan-Jun 2023
                01 March 2023
                : 17
                : 1
                : 53-57
                Affiliations
                [1] Department of Transfusion Medicine, Sanjay Gandhi Post Graduate Institute of Medical Science, Lucknow, Uttar Pradesh, India
                [1 ] Department of Immunohematology and Blood Transfusion, Kalyan Singh Super Speciality Cancer Institute, Lucknow, Uttar Pradesh, India
                [2 ] Department of Transfusion Medicine, S.N. Medical College, Agra, Uttar Pradesh, India
                Author notes
                Address for correspondence: Dr. Anju Dubey, Department of Immunohematology and Blood Transfusion, Kalyan Singh Super Speciality Cancer Institute, Lucknow, Uttar Pradesh, India. E-mail: dranjudubey@ 123456gmail.com
                Article
                AJTS-17-53
                10.4103/ajts.ajts_115_22
                10180786
                37188031
                a96d81bb-3801-45b0-820d-62a731017d85
                Copyright: © 2023 Asian Journal of Transfusion Science

                This is an open access journal, and articles are distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as appropriate credit is given and the new creations are licensed under the identical terms.

                History
                : 07 September 2022
                : 11 October 2022
                : 06 November 2022
                Categories
                Original Article

                Hematology
                alloimmunization,genotype,phenotype,thalassemia
                Hematology
                alloimmunization, genotype, phenotype, thalassemia

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