Design and development of a PCR method for detection of SNP of CYP2B6 / Tengku Aliaa Dayana Tuan Mohd Hashim

Cytochrome P450 286 is an enzyme that involved in the biotransformation of many clinically important drugs. Genetic polymorphisms of the CYP2B6 gene are an important factor that contributes to the inter-individual and inter-ethnic variability in the systemic exposure, therapeutic and toxic responses to CYP2B6 substrate drugs. Therefore, it is important to develop a method to identify and determine all of the genetic variants in a CYP2B6 gene that responsible for altered CYP2B6 expression. The aim of this study was to design and develop rapid and simple method for the detection of SNP in the CYP2B6 gene. The multiplex, allele-specific PCR method was used. This method involves two-stage procedures, which were first and second PCR. First PCR was used to amplify 3 exons, which were exon 4, exon 5+6, and exon 9. After that, the products of the first PCR were used as a DNA template and genotyped by allele-specific PCR. The assay detected the following published single nucleotide polymorphisms such as G516T (Gln172His), A785G (Lys262Arg) and C1459T (Arg487Cys). DNA sequencing analysis was performed to further confirmed genotype results obtained from current method. Multiplex PCR with allele-specific genotyping method is simple, reliable, rapid and maybe more economical compared to other methods to perform. Further studies are required in order to investigate the impact of the SNP of CYP2B6 gene on the clinical response to drugs that are substrates for this gene.