Ludmila Alves Sanches Dutra Patrícia Godoy Garcia Costa Lara Franciele Ribeiro Velasco Angélica Amorim Amato

Objective: The aim of the present study is to validate a rapid and simple allelespecifi c 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 specifi c for one of the two variants. They were combined with a common reverse primer into two PCR reactions. Specifi c amplifi cation 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-specifi c 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 identifi cation. (Arq Bras Endocrinol Metab 2008; 52/8:1362-1366)