Comparison of TaqMan Real-Time and Tetra-Primer ARMS PCR Techniques for Genotyping of Rs 8066560 Variant in Children and Adolescents with Metabolic Syndrome.

Abstract

Single nucleotide polymorphisms (SNPs) are major contributors to susceptibility or resistance to various human diseases. Metabolic syndrome (MetS) is a collection of risk factors, including abdominal obesity, dyslipidemia and increased blood pressure. MetS is more common among Iranian children and adolescents in comparison to other ethnicities. Sterol regulatory element binding factors (SREBFs) involve in the regulation of carbohydrate and lipid metabolism. The tetra-primer amplification refractory mutation system PCR (tetra-primer ARMS-PCR/T-ARMS-PCR) is a fast and economical means of assaying SNPs, requiring only PCR amplification and subsequent electrophoresis for the determination of genotypes. This study aims to optimize and compare the results of tetra-primer ARMS-PCR for genotyping of rs8066560 in Iranian children and adolescents being afflicted with metabolic syndrome with the TaqMan assay. In this study, a total of 50 individuals with 9-19 years of age, including 25 healthy subjects and 25 MetS cases were studied. The tetra-primer ARMS-PCR was used to genotype the rs8066560. After PCR optimization, we could successfully detect the rs8066560 polymorphism in all the studied subjects. Furthermore, we observed complete concordance between tetra-primer ARMS-PCR assay and TaqMan method`s results. Tetra-primer ARMS-PCR can be utilized as a cost-effective, rapid and reproducible method for SNP genotyping especially while performing large-scale epidemiological/association at studies.