The development of an allele­-specific polymerase chain reaction (AS-PCR) based genotyping for CYP2D6*10 allele

CYP2D6 is a drug metabolising enzyme that metabolises a vast majority of drugs including psychotropic, beta blockers, and antidepressants. Its gene is associated with single nucleotide polymorphism (SNP) and one of its variants is CYP2D6*10 which is associated with the decrease in the CYP2D6 activity. This variant is mostly found in Asian and it is particularly important as it can be used to determine the responses of individuals towards drug metabolism. For this reason, an allele-specific polymerase chain reaction (AS-PCR) method was developed to provide a simple and validated cost-effective PCR based technique to screen for CYP2D6* 10 in human DNA samples. In this method, allele-specific primers were designed to have the same 3 '-end as the SNP so that it would complement the DNA template at particular SNP site during PCR amplification. The amplicons were then separated by using agarose gel electrophoresis. Four samples were tested for their genotype and direct sequencing was used to validate this method. This method provides a simple and cost-effective genotyping way to study CYP2D6* 10 using conventional instrumentation.