Molecular typing for the Indian blood group associated 252G>C single nucleotide polymorphism in a selected cohort of Australian blood donors.
| Autor: | Lopez GH; Research and Development Division, Australian Red Cross Blood Service, Queensland, Australia., Mcbean RS; Research and Development Division, Australian Red Cross Blood Service, Queensland, Australia., Wilson B; Red Cell Reference Laboratory, Australian Red Cross Blood Service, Queensland, Australia., Irwin DL; Sequenom Inc. Asia Pacific, Herston, Queensland, Australia., Liew YW; Red Cell Reference Laboratory, Australian Red Cross Blood Service, Queensland, Australia., Hyland CA; Research and Development Division, Australian Red Cross Blood Service, Queensland, Australia., Flower RL; Research and Development Division, Australian Red Cross Blood Service, Queensland, Australia. |
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| Jazyk: | angličtina |
| Zdroj: | Blood transfusion = Trasfusione del sangue [Blood Transfus] 2015 Jan; Vol. 13 (1), pp. 78-85. Date of Electronic Publication: 2014 Jun 05. |
| DOI: | 10.2450/2014.0336-13 |
| Abstrakt: | Background: The Indian blood group antigens, In(a) and In(b), are clinically significant in transfusion medicine. However, antisera to type these antigens are difficult to obtain. The In(b) antigen is a high frequency antigen present in all populations, while the frequency of the antithetical In(a) ranges from 0.1% in Caucasians up to 11% in Middle Eastern groups. This antigen polymorphism is encoded by the single nucleotide polymorphism (SNP) 252G>C in CD44. The aim of this study was to establish and compare two genotyping methods to measure the frequency of the IN*A and IN*B alleles in a blood donor cohort. Materials and Methods: Donor blood samples (n=151) were genotyped by a novel real-time polymerase chain reaction (PCR) high-resolution meltcurve (HRM) analysis and a custom matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) assay. Samples with the rare IN*A allele were further investigated by nucleotide sequencing, red cell agglutination, and flow cytometry techniques. Results: In this study group, 149 IN*B homozygous and 2 IN*A/B heterozygous samples were detected with 100% concordance between HRM and MALDI-TOF MS methods. For PCR HRM, amplicon melting alone did not differentiate IN*A and IN*B alleles (class 3 SNP), however, the introduction of an unlabelled probe (UP) increased the resolution of the assay. Sequencing confirmed that the two non-homozygous samples were IN*A/B heterozygous and phenotyping by red cell agglutination, and flow cytometry confirmed both In(a) and In(b) antigens were present as predicted. Discussion: Genotyping permits conservation of rare antisera to predict blood group antigen phenotype. In PCR UP-HRM the IN*A and IN*B alleles were discriminated on the basis of their melting properties. The In(a) frequency in this selected donor population was 1.3%. Application of genotyping methods such as these assists in identifying donors with rare blood group phenotypes of potential clinical significance. |
| Databáze: | MEDLINE |
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