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. 2018 Dec;148(6):713-720.
doi: 10.4103/ijmr.IJMR_455_17.

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

Swati Kulkarni  1 , Bhavika Choudhary  1 , Harita Gogri  1 , Shashikant Patil  2 , Mamta Manglani  3 , Ratna Sharma  3 , Manisha Madkaikar  1
Affiliations

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

Swati Kulkarni et al. Indian J Med Res. 2018 Dec.

Abstract

Background & objectives: In multitransfused thalassaemic patients, haemagglutination fails to phenotype the patient's blood group antigens due to the presence of donor-derived erythrocytes. DNA-based methods can overcome the limitations of haemagglutination and can be used to determine the correct antigen profile of these patients. This will facilitate the procurement of antigen-matched blood for transfusion to multitransfused patients. Thus, the aim of this study was to compare the serological phenotyping of common and clinically important antigens of Rh, Duffy, Kell, Kidd and MNS blood group systems with molecular genotyping amongst multitransfused thalassaemic patients.

Methods: Blood samples from 200 patients with thalassaemia and 100 'O' group regular blood donors were tested using standard serological techniques and polymerase chain reaction-based methods for common antigens/alleles (C, c, D, E, e, Fya, Fyb, Jka, Jkb, K, k, M, N, S, s).

Results: Genotyping and phenotyping results were discordant in 77 per cent of thalassaemic patients for five pairs of antithetical antigens of Rh, Duffy, Kell and Kidd blood group systems. In the MNS blood group system, 59.1 per cent of patients showed discrepancy. The rate of alloimmunization among thalassaemics was 7.5 per cent.

Interpretation & conclusions: Molecular genotyping enabled the determination of the actual antigen profile in multitransfused thalassaemia patients. This would help reduce the problem of alloimmunization in such patients and would also aid in the better management of transfusion therapy.

Keywords: Blood group genotyping; blood group systems; haemagglutination; molecular genotyping; polymerase chain reaction-sequence-specific primer; thalassaemia.

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Conflict of interest statement

None

Figures

Figure 1
Figure 1
Standardization of RBC antigen genotyping by PCR-SSP (A) FY1, FY2, JK1, JK2, K1 and K2 alleles; (B) C, c, D, E, and e antigens; (C) M, N, S and s antigens: M1, M2, M3 and M4 represent amplification of different regions of GYPA responsible for M antigen specificity (as followed by Heymann and Salama, 2010). Presence of all the specific fragments accounts for the presence of M antigen. Similarly, N1 and N2 represent amplification of two regions of GYPA responsible for N antigen specificity. S/s polymorphism is due to a SNP, hence a single PCR each for S and s has been used for genotyping. PCR, polymerase chain reaction; SSP, sequence-specific primer; SNP, single-nucleotide polymorphism; RBC, red blood cell.
Figure 2
Figure 2
Percentage discrepancy of clinically important blood group antigens by serotyping and genotyping among thalassaemia patients.
See this image and copyright information in PMC

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