Khatami Mehri, Heidari Mohammad Mehdi, Soheilyfar Sorour
Department of Biology, Faculty of Science, Yazd University, Yazd, Iran.
Department of Biology, Ashkezar Islamic Azad University, Ashkezar, Yazd, Iran.
Arch Med Sci Atheroscler Dis. 2016 Apr 27;1(1):e9-e15. doi: 10.5114/amsad.2016.59587. eCollection 2016.
The T to C transition at nucleotide 1565 of the human glycoprotein IIIa () gene represents a genetic polymorphism (PlA1/A2) that can influence both platelet activation and aggregation and that has been associated with many types of disease. Here, we present a newly designed multiplex tetra-primer amplification refractory mutation system - polymerase chain reaction (T-ARMS-PCR) for genotyping a single nucleotide polymorphism (SNP) (dbSNP ID: rs5918) in the human gene.
We set up T-ARMS-PCR for the rs5918 SNP in a single-step PCR and the results were validated by the PCR-RFLP method in 132 coronary artery disease (CAD) patients and 122 unrelated healthy individuals.
Full accordance was found for genotype determination by the PCR-RFLP method. The multiple logistic regression analysis showed a significant association of the rs5918 polymorphism and CAD according to dominant and recessive models (dominant model OR: 2.40, 95% CI: 1.33-4.35; = 0.003, recessive model OR: 4.71, 95% CI: 1.32-16.80; = 0.0067).
Our T-ARMS-PCR in comparison with RFLP and allele-specific PCR is more advantageous because this PCR method allows the evaluation of both the wild type and the mutant allele in the same tube. Our results suggest that the rs5918 (PlA1/A2) polymorphism in the gene may contribute to the susceptibility of sporadic Iranian coronary artery disease (CAD) patients.
人类糖蛋白IIIa（）基因第1565位核苷酸处的T到C转换代表一种基因多态性（PlA1/A2），它可影响血小板活化和聚集，并与多种疾病相关。在此，我们提出一种新设计的多重四引物扩增阻滞突变系统-聚合酶链反应（T-ARMS-PCR），用于对人类基因中的单核苷酸多态性（SNP）（dbSNP ID：rs5918）进行基因分型。
我们在一步PCR中建立了针对rs5918 SNP的T-ARMS-PCR，并通过PCR-RFLP方法在132例冠状动脉疾病（CAD）患者和122名无关健康个体中验证了结果。
通过PCR-RFLP方法进行基因型测定时发现完全一致。多元逻辑回归分析显示，根据显性和隐性模型，rs5918多态性与CAD存在显著关联（显性模型OR：2.40，95%CI：1.33-4.35；=0.003，隐性模型OR：4.71，95%CI：1.32-16.80；=0.0067）。
与RFLP和等位基因特异性PCR相比，我们的T-ARMS-PCR更具优势，因为这种PCR方法允许在同一管中评估野生型和突变等位基因。我们的结果表明，基因中的rs5918（PlA1/A2）多态性可能导致散发性伊朗冠状动脉疾病（CAD）患者易感性增加。
