Development of One-Step Tetra-primer ARMS-PCR for Simultaneous Detection of the I/D and Gene Polymorphisms and the Correlation with CAD Patients.

Background: 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.

Methods: 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.

Results: 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.

Conclusion: 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.