Mutation detection in human estrogen receptor B gene in infertile male patients by denaturing high - performance liquid chromatography

For screening sequence variations in genes, rapid turnover time is of fundamentalimportance. While, many of the current methods are unfortunately time consuming and technicallydifficult to implement. Denaturing high-performance liquid chromatography (DHPLC) method hadbeen shown to be a high-throughput, time saving, and economical tool for mutation screening.

In the present study DHPLC method was used to explore the potential associationbetween estrogen receptor β gene (ESR2) variants and male infertility.

DNA from 96 men with infertility and 96 normal male as control werescreened for mutation in the nine exons of the ESR2 gene, using WAVE® DHPLC device equippedwith a DNA separation column and automated sequence analysis on the ABI Prism 310.

DHPLC evaluation of ESR2 gene in 96 infertile patients, revealed one heterozygoussequence variation (IVS 8–4G>A) near the 5’ splicing region of intron 8 in 5 patients. No variationwas identified in control population.

Mutation detection by DHPLC, as it is presented in this context, is a high-throughput,quick, and economical tool for mutation screening. The gene alterations in ESR2 gene that we’vefound might increase susceptibility to infertility; but without cDNA screening, the consequences ofthese genetic alterations cannot be predicted.