Rapid genotyping of 508G>A (rs3745635) and 1067T>A (rs3894326) of FUT3 by a duplex Eprobe‐mediated melting curve analysis

Lewis histo-blood group phenotypes are regulated by the action of FUT3-encoded α(1,3/1,4)fucosyltransferase and FUT2-encoded α(1,2)fucosyltransferase. Since Lewis phenotypes are suggested to be associated with various clinical conditions, a method for large-scale FUT3 genotyping is desirable. In worldwide populations, 508G>A and 1067T>A of FUT3 are two of three common causal single nucleotide polymorphisms for Lewis-negative alleles.We developed a duplex Eprobe-mediated melting curve analysis for genotyping 508G>A and 1067T>A simultaneously and applied this method to 106 Ghanaian and 140 Japanese subjects.The results of both 508G>A and 1067T>A genotyping by duplex Eprobe-mediated melting curve analysis were completely in agreement with the results of a DNA sequence analysis in 106 Ghanaians and polymerase chain reaction-restriction fragment length polymorphism analysis in 140 Japanese subjects.The present duplex Eprobe-mediated melting curve analysis is valid and credible for large-scale estimation of Lewis-negative alleles.