Development of Cost-effective Tetra-primer Amplification Refractory Mutation System (T-ARMS) PCR for the Detection of miR-146a gene rs2910164 C/G Polymorphism in Breast Cancer

Muhammad Afzal, Amena Rahim, A

Abstract

Evaluation of a single nucleotide polymorphism (SNP) and single nucleotide mutation in cancer patients and other diseases is crucial for genotypic characterization in order to select therapy and genetic counseling. The study aim is to develop and optimization for the detection of miR-146a gene rs2910164 C/G polymorphism in breast cancer. Tetra-primer Amplification Refractory Mutation System (T-ARMS) PCR was applied on 60 individuals to determine the genotype and allele frequencies. Genotype frequencies of miR-146a gene rs2910164 C/G polymorphism were homozygous CC 65%; heterozygous CG 6.65%; homozygous GG 28.35%; allele C and G frequencies were 68.5% and 31.5% respectively. These were in Hardy–Weinberg’s equilibrium; p=0.18. Our study provides a simple rapid and a cost-effective T-ARMS-PCR molecular technique that could be used for the detection allele specific DNA polymorphisms and mutations in certain diseases and normal population at large scale.