Establishment of an alternative efficiently genotyping strategy for human ABO gene

Abstract

ABO genotyping is used in several disciplines, including transfusion, transplantation, human evolution, and forensic medicine. Detection of single nucleotide polymorphisms (SNPs) on a locus is a common way to identify different genotypes. In this study we developed a strategy for ABO genotyping, which can rapidly and efficiently detect SNPs. DNA fragments containing 4 SNPs in the ABO gene (c.261delG, c.297A > G, c.1009A > G, and c.1061delC) were amplified using individually and multiplexed polymerase chain reaction (PCR)-based methods and subsequently genotyped by high-resolution melting (HRM) analysis. Human blood ABO genotypes from 92 samples were successfully determined by HRM analysis. A total of 14 genotypes (A/A, A/O01, A/O02, A201/O01, A205/O01, B/B, B/O01, B/O02, A/B, A201/B, A205/B, O01/O01, O02/O02, O01/O02) were identified by analysis of the 4 SNPs of interest in this study. The results suggest that the present HRM assay is a reliable and rapid method for ABO blood type genotyping and it may offer an alternative to traditional genotyping methods.

Keywords: ABO genotyping; Fluorescence melting curve analysis; HRM; Single nucleotide polymorphism.