Detection of c.375A>G, c.385A>T, c.571C>T, and of via Real-Time PCR in a Single Tube.

α(1,2)fucosyltransferase (Se enzyme) encoded by is involved in the secretor status of ABH(O) blood group antigens. The allele is one of the non-functional () alleles in which 9.3 kb, containing the entire coding region of , is deleted by -mediated nonhomologous recombination. In addition to this allele, three SNPs of , c.375A>G, c.385A>T, and c.571C>T, appear to be prevalent in certain Oceanian populations such as Polynesians. Recently, we developed an endpoint genotyping assay to determine zygosity, using a FAM-labeled probe for detection of the allele and a VIC-labeled probe for the detection of . In this study, instead of the VIC probe, a HEX-labeled probe covering both c.375A>G and c.385A>T and a Cy5-labeled probe covering c.571C>T were added to the allele assay mixture to allow for the simultaneous detection of these four variations via endpoint genotyping for zygosity and fluorescence melting curve analysis for c.375A>G, c.385A>T, and c.571C>T genotyping. The results obtained from 24 Samoan subjects using this method were identical to those obtained using previous methods. Therefore, it appears that the present method can accurately determine these four variations simultaneously.