Designing and Validation of One-Step T-ARMS-PCR for Genotyping the eNOS rs1799983 SNP

Abstract

<strong>Background:</strong> The transversion of G to T (G894T) in human endothelial nitric oxide synthase (eNOS) gene has profound effects such as male infertility, recurrent miscarriage, multiple sclerosis and cardiovascular diseases.<br />Objectives: Development of a new Multiplex Tetra-Primer Amplifi cation Refractory Mutation System - Polymerase Chain Reaction (T-ARMS-PCR) for detection of rs1799983 (G894T) in the human eNOS was sought.<br /><strong>Materials and Methods:</strong> A T-ARMS-PCR for rs1799983 polymorphism in a single-step PCR was carried out, and the results were confi rmed by PCR-RFLP technique in 82 infertile men with varicocele.<br /><strong>Results:</strong> The results showed that GG (varicocele infertile men), GT and TT genotypes appear to be 53.65%, 34.14%, and 12.19%, respectively. Full accordance between PCR-RFLP and T-ARMS-PCR methods for genotyping of rs1799983 polymorphism was found.<br /><strong>Conclusions:</strong> This is the fi rst work that describes a rapid, relatively cheap, high throughput detection of G894T polymorphism in eNOS that can be used in large scale clinical studies.