Allele-specific PCR assay to genotype SNP rs7903146 in TCF7L2 gene for rapid screening of diabetes susceptibility

Abstract

Objective: The aim of the present study is to validate a rapid and simple allele-specific PCR that genotypes TCF7/L2 rs7903146 (C/T) polymorphism with standard PCR instruments. Methods: Two forward primers with variations in their 3′ nucleotides were designed in such a way that each was specific for one of the two variants. They were combined with a common reverse primer into two PCR reactions. Specific amplification indicates the presence of the allele. One hundred and four DNA samples were genotyped by this method. To evaluate the assay, the polymorphism spanning region of 63 DNA samples representing the three possible genotypes was sequenced. Results: The rs7903146 allele assignments derived from the allele-specific PCR were in complete agreement with sequencing. Conclusions: The assay described here is a suitable strategy for the TCF7/L2 rs7903146 (C/T) genotyping also allowing rapid and reliable identification.