Optimisation of the Tetra primer ARMS PCR protocol for detecting the single-nucleotide polymorphism rs1799929 of the NAT2 gene in humans from some provinces, cities in Vietnam

Abstract

The single-nucleotide polymorphism (SNP) rs1799929 in the NAT2 gene has been identified as being associated with drug metabolism and the risk of some serious diseases. This study developed a protocol to detect the SNP rs1799929 in the NAT2 gene using the Tetra primer - Amplification refractory mutation system polymerase chain reaction (T-ARMS PCR) technique. The objective of the study was to optimise and apply this technique for the detection of the NAT2 rs1799929 SNP. We employed the T-ARMS PCR technique to amplify the gene region containing the
SNP and compared the results with the Sanger sequencing method for validation. The study demonstrated that the protocol was successfully optimised, including adjustments of annealing temperature and primer concentrations to ensure high accuracy and specificity. Notably, the SNP detection by T-ARMS PCR showed 100% concordance with the Sanger sequencing method, confirming the reliability of this technique. The results of this study provide a foundation for application in further studies with larger sample sizes.