DEVELOPMENT OF A METHOD FOR GENOTYPING THE 𝘔𝘛𝘏𝘍𝘙 RS1801133 POLYMORPHISM IN VIETNAMESE, USING THE POLYMERASE CHAIN REACTION WITH CONFRONTING TWO-PAIR PRIMERS

Abstract

Aims: To develop a method using polymerase chain reaction with confronting two-pair primers (PCR-CTPP) to genotype MTHFR rs1801133 polymorphism in Vietnamese people. Methods: DNA samples from 15 Vietnamese people were used to evaluate this assay. The Sanger sequencing method was used to identify the standard genotypes of the rs1801133 polymorphism. Some bioinformatic softwares were used to design four a multiplex PCR-CTPP. Experimental procedures were conducted to verify the primers and components of the PCR method. Results: The protocol of the PCR-CTPP was successful developed with optimal inner/outer primers ratio, the melting temperature of primers, the components and the thermal cycling to identify MTHFR rs1801133 polymorphism. Conclusion: The PCR-CTPP protocol should be applied to genotyping MTHFR rs1801133 polymorphism in large cohorts to investigate the association between the polymorphism and metabolic syndrome in Vietnamese population.