Molecular genotyping of clinically important blood group antigens in patients with thalassaemia

Kulkarni, Shrikant; Choudhary, Bhavika; Gogri, Harita; Patil, Shashikant; Manglani, Mamta; Sharma, Rachana; Madkaikar, Manisha

doi:10.4103/ijmr.ijmr_455_17

Cited by 24 publications

(15 citation statements)

References 28 publications

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“…Donors were typed for common blood group antigens of Rh (C, c, D, E, and e), Duffy (Fy a and Fy b ), Kell (K and k) and Kidd (Jk a and Jk b ) using commercially available antiserum using the manufacturer's instructions (IMMUCOR, Inc., USA) by conventional tube technique. The antigen profiles were analyzed for providing antigen-matched blood to 84 non-alloimmunized and 15 alloimmunized, multitransfused thalassaemia major patients from our previous study 8 . Antigen profiles of these patients were determined by PCR using sequence-specific primer (PCR-SSP) and haemagglutination.…”

Section: Methodsmentioning

confidence: 99%

“…For PCR, DNA was prepared from ethylenediaminetetraacetic acid (EDTA) blood using commercially available DNA extraction kit (Qiagen, Germany). The common alleles of Rh, Duffy, Kell, Kidd and MNS antigens were genotyped using PCR-SSP along with known controls in Veriti ® 96-well Thermal Cycler (Applied Biosystems, USA) as described earlier 8 . The amplified products were separated electrophoretically on two per cent agarose gel containing ethidium bromide and visualized under ultraviolet transilluminator, Gel Doc system (Bio-Rad, USA).…”

Section: Methodsmentioning

confidence: 99%

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Red cell antigen phenotypes in blood donors & thalassaemia patients for creation of red cell antigen-matched inventory

Kulkarni

Choudhary

Gogri

et al. 2020

Indian Journal of Medical Research

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Background & objectives : Patients with thalasssaemia are at a risk of alloimmunization and the presence of RBC alloantibodies further complicates transfusion therapy. Matching for the critical antigens of Rh, Kell, Kidd and Duffy blood group systems has been shown to minimize alloimmunization. The aim of the present study was to create a database of extensively typed donors for clinically significant and common blood group antigens of Rh, Kidd, Kell and Duffy systems for transfusion therapy of multitransfused thalassaemic patients. Methods : Five hundred O group regular blood donors were phenotyped for Rh, Kell, Duffy and Kidd blood group antigens using haemagglutination technique. Eighty four non-alloimmunized and 15 alloimmunized thalassaemia major patients with known antigenic profiles (determined by polymerase chain reaction with sequence-specific primers) were selected for this study. Results : By analyzing antigen profiles of 500 O group regular donors, a database of 193 donors matching perfectly for Rh, Duffy, Kell and Kidd antigens was prepared for 15 alloimmunized patients. For non-alloimmunized 84 thalassaemic patients, a database of 405 donors was created. Interpretation & conclusions : A database of 500 regular blood donors phenotyped for common antigens of Rh, Duffy, Kell and Kidd blood group systems was created, which would be useful in providing extended antigen-matched RBCs for thalassaemia patients. This will improve the quality and effectiveness of transfusion therapy.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Red cell antigen phenotypes in blood donors & thalassaemia patients for creation of red cell antigen-matched inventory

Kulkarni

Choudhary

Gogri

et al. 2020

Indian Journal of Medical Research

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“…20,21 In part, they make it possible to overcome the limitations of serological tests. [22][23][24][25] In immunohematology, molecular methods have become more popular since the 1990s when the genetic background of antigen specificities was first described. 26,27 A single nucleotide variant (SNV) is the most common variation resulting from a change of amino acid in the protein sequence which leads to either the presence or absence of the antigen.…”

Section: Blood Group Genotypingmentioning

confidence: 99%

“…Study results support extended profiling of donors and patients for the best prophylactic antigen matching to prevent alloimmunization. 16,[22][23][24] Literature on the use of NGS for studying blood group antigens shows a picture of rapidly developing technology which may prove highly reliable. The technology is now being validated and upgraded especially in terms of antigens with a complex genetic background or with highly homological regions.…”

Section: Ngs In Blood Group Screeningmentioning

confidence: 99%

<p>Potential of next-generation sequencing to match blood group antigens for transfusion</p>

Orzińska¹,

Guz²,

Brojer³

2019

IJCTM

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A review of the advances in applying next-generation sequencing (NGS) to transfusion medicine for the purpose of genotyping alleles encoding clinically important red blood cell and platelet antigens. NGS data from published studies confirm the possibility of antigen prediction based on sequencing of the whole genome, exome or targeted regions. What remains a challenge, to provide highly accurate NGS genotyping, is the further improvement of bioinformatic solutions for automated interpretation based on publicly accessible and improved reference databases appropriate for NGS methods as well as validation of a method based on the examination of a large number of individuals. There is no doubt, however, as to the future of NGS as a supplementary test used to provide highly compatible blood as well as to reduce the risk of patient's alloimmunization. This is part of personalized medicine.

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“…The polymerase chain reaction (PCR) is a vital tool for erythrocyte genotyping, moreover PCR and gel electrophoresis are easy, low cost, and flexible methods for amplifying specific target DNA. Allele Specific PCR (AS-PCR), Multiplex-PCR, and Restriction Fragment Length Polymorphism-PCR (RFLP-PCR) techniques are commonly used to identify erythrocyte polymorphisms [ 8 – 10 ].…”

Section: Introductionmentioning

confidence: 99%

Genotyping versus phenotyping of non-ABO erythrocyte antigens in patients with the Mediterranean hemopathic syndromes: Effect of transfusion therapy

et al. 2021

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The Mediterranean hemopathic syndromes (MHS) are the most prevalent hemoglobinopathies in the Mediterranean basin. Transfusion therapy is the main therapy for these disorders, particularly for severe forms of the disease. Currently, pre-transfusion serological typing of erythrocyte antigens is the standard tool for reducing complications of transfusion in those patients. This study compared genotyping with phenotyping of non-ABO erythrocyte antigens in patients with MHS and assessed the effect of transfusion therapy on their results. One-hundred ninety-eight MHS patients were recruited, screened, and proven negative for allo-antibodies. They were grouped into two groups: (1) 20 newly diagnosed patients with no transfusion history and (2) 178 previously diagnosed patients undergoing transfusion therapy. Patients were interviewed and clinically examined. Full blood count (FBC) and high performance liquid chromatography (HPLC) were done for group 1 only. Genotyping and phenotyping of non-ABO erythrocyte antigens were performed for group 1, and 25 patients out of group 2 were propensity score-matched (PSM) with group 1. Both groups were gender and age matched; 55% and 74% of groups 1 and 2 had major disease, respectively. Insignificant differences were observed between genotyping and phenotyping of non-ABO erythrocyte antigens in group 1, while significant discrepancies and mixed field results were noted in group 2 patients. Discrepancies were obvious with JKa, JKb, and little c antigens. Conclusively, molecular typing is a powerful tool for pre-transfusion testing in chronically transfused MHS patients. This testing reduces incidence of transfusion reactions. JKa, JKb and little c antigens are the most clinically significant non-ABO erythrocyte antigens.

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Molecular genotyping of clinically important blood group antigens in patients with thalassaemia

Cited by 24 publications

References 28 publications

Red cell antigen phenotypes in blood donors & thalassaemia patients for creation of red cell antigen-matched inventory

Red cell antigen phenotypes in blood donors & thalassaemia patients for creation of red cell antigen-matched inventory

<p>Potential of next-generation sequencing to match blood group antigens for transfusion</p>

Genotyping versus phenotyping of non-ABO erythrocyte antigens in patients with the Mediterranean hemopathic syndromes: Effect of transfusion therapy

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