Heidar Mohammad Mehdi, Khatami Mehri
Department of Biology, Faculty of Science, Yazd University, Yazd, Iran.
Iran J Biotechnol. 2017 Sep 27;15(3):208-212. doi: 10.15171/ijb.1307. eCollection 2017.
The transversion of G to T (G894T) in human endothelial nitric oxide synthase () gene has profound effects such as male infertility, recurrent miscarriage, multiple sclerosis and cardiovascular diseases. Development of a new Multiplex Tetra-Primer Amplification Refractory Mutation System - Polymerase Chain Reaction (T-ARMS-PCR) for detection of rs1799983 (G894T) in the human was sought. A T-ARMS-PCR for rs1799983 polymorphism in a single-step PCR was carried out, and the results were confirmed by PCR-RFLP technique in 82 infertile men with varicocele. 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. This is the first work that describes a rapid, relatively cheap, high throughput detection of G894T polymorphism in that can be used in large scale clinical studies.
人类内皮型一氧化氮合酶（）基因中G到T的颠换（G894T）具有深远影响，如男性不育、复发性流产、多发性硬化症和心血管疾病。人们试图开发一种新的多重四引物扩增难治性突变系统 - 聚合酶链反应（T-ARMS-PCR）来检测人类中的rs1799983（G894T）。对82名精索静脉曲张不育男性进行了一步法PCR检测rs1799983多态性的T-ARMS-PCR，并通过PCR-RFLP技术对结果进行了验证。结果显示，GG（精索静脉曲张不育男性）、GT和TT基因型分别占53.65%、34.14%和12.19%。发现rs1799983多态性基因分型的PCR-RFLP和T-ARMS-PCR方法完全一致。这是首次描述一种快速、相对廉价、高通量检测中的G894T多态性的方法，可用于大规模临床研究。
