Tetra-primer Amplification Refractory Mutation System Polymorphism Chain Reaction for Genotyping of Dyslipidemia-associated Alleles of CELSR2, APOB, and APOE Genes

Abstract Background: Dyslipidemia refers to an abnormal level of plasma lipids, mainly high levels of low-density lipoproteins and triglycerides, and low levels of high-density lipoproteins. This lipid abnormality is demonstrated to be strongly linked to the development of cardiovascular diseases. Genome-wide association studies have shown several single nucleotide polymorphisms (SNPs) linked with dyslipidemia. Materials and Methods: In this investigation, we have optimized a tetra-primer amplification refractory mutation system (T-ARMS) polymorphism chain reaction (PCR) protocol for four SNPs, i.e., CELSR2 (rs629301), APOB (rs1367117), APOE (rs429358), and APOE (rs7412) associated with dyslipidemia. Results: T-ARMS PCR is a multiplex, sensitive, and low-technique to genotype the SNPs performed in a single tube and reaction. Several factors, such as DNA purity, annealing temperature, PCR reagent equilibrium, and concentration of magnesium chloride in the reaction, influence the assay. However, the T-ARMS PCR assay has been demonstrated to be unsuitable for variants rich in guanine and cytosine. Conclusion: T-ARMS PCR is the method of choice for association studies of variants with genetic diseases and provides a fast, accurate, reliable, and low-cost way to study SNPs.