A primer extension denaturing high-performance liquid chromatography method for the identification of three ABCC2 genetic polymorphisms

Abstract

Background: A number of single nucleotide polymorphisms have been described in the ABCC2 gene that alters drug disposition. The aim of the study was to develop a primer extension denaturing high-performance liquid chromatography (PE-dHPLC) assay to determine three common variants of the ABCC2 gene in the promoter region (−24C>T), exon 10 (1249G>A), and exon 28 (3972C>T). Methods: Polymerase chain reactions (PCRs) were used to isolate the area of interest in the ABCC2 gene. A comparison of the PCR product was performed between sequencing and dHPLC. Results: A 100% identity match was achieved between groups and allowed for a quick and accurate method to determine three single nucleotide polymorphisms in a single extension reaction. This assay is the first of its type to determine three ABCC2 variants by dHPLC