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

ABSTRACT

For screening sequence variations in genes, rapid turnover time is of fundamental importance. While, many of the current methods are unfortunately time consuming and technically difficult to implement. Denaturing high-performance liquid chromatography [DHPLC] method had been 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 association between estrogen receptor gene [ESR2] variants and male infertility. DNA from 96 men with infertility and 96 normal male as control were screened for mutation in the nine exons of the ESR2 gene, using WAVE [R] DHPLC device equipped with a DNA separation column and automated sequence analysis on the ABI Prism 310. DHPLC evaluation of ESR2 gene in 96 infertile patients, revealed one heterozygous sequence variation [IVS 8-4G>A] near the 5' splicing region of intron 8 in 5 patients. No variation was 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've found might increase susceptibility to infertility; but without cDNA screening, the consequences of these genetic alterations cannot be predicted