Heidari Mohammad Mehdi, Hadadzadeh Mehdi, Fallahzadeh Hossein
Department of Biology, Faculty of Science, Yazd University, Yazd, Iran.
Department of Cardiac Surgery, Afshar Hospital, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
Avicenna J Med Biotechnol. 2019 Jan-Mar;11(1):118-123.
The Insertion/Deletion and gene polymorphisms could be associated with pathogenesis of essential hypertension and cardiovascular disorders and Coronary Artery Disease (CAD). In the present study, a fast and novel approach of multiplex Tetra-Primer Amplification Refractory Mutation System-PCR (T-ARMS-PCR) was developed for simultaneous detection of two SNPs including (rs4340) and 2350A>G (rs4343) of () gene.
The present research was performed using 148 blood samples taken from patients with CAD and 135 healthy individuals. One set of inner primers (for rs4343) and one set of outer primer pairs were designed for genotyping of Insertion/Deletion and rs4343 polymorphisms in single tube T-ARMS-PCR.
Our results manifested that genotypes and alleles frequency of the polymorphisms showed no statistically significant association between CAD patients and the control group. In addition, complete concordance was seen between sensitive Tetra-ARMS-PCR and sequencing method.
The technique is the first work for simultaneous detection of Insertion/Deletion polymorphism and rs4343 SNPs in gene and the results were entirely according to those from an independent procedure.
插入/缺失及基因多态性可能与原发性高血压、心血管疾病及冠状动脉疾病（CAD）的发病机制相关。在本研究中，开发了一种快速且新颖的多重四引物扩增阻滞突变系统聚合酶链反应（T-ARMS-PCR）方法，用于同时检测（）基因的两个单核苷酸多态性（SNP），即（rs4340）和2350A>G（rs4343）。
本研究使用从CAD患者采集的148份血样和135名健康个体进行。设计了一组内引物（用于rs4343）和一组外引物对，用于在单管T-ARMS-PCR中对插入/缺失及rs4343多态性进行基因分型。
我们的结果表明，多态性的基因型和等位基因频率在CAD患者和对照组之间无统计学显著关联。此外，灵敏的四引物扩增阻滞突变系统聚合酶链反应（Tetra-ARMS-PCR）与测序方法之间具有完全一致性。
该技术是首次同时检测基因中的插入/缺失多态性和rs4343单核苷酸多态性，结果与独立检测程序的结果完全一致。
