Fast and more affordable CYP2C19 LOF testing to personalized of clopidogrel therapy

Abstract Background Clopidogrel is an antiplatelet drug used to treat coronary artery disease (CAD) reduce blood clotting by inhibiting platelet activity. However, many patients experience ischemic events due changes in clopidogrel metabolism attributed various genetic and non-genetic factors. CYP2C19 one of the important genes responsible for clopidogrel, but this gene has polymorphisms cytochrome P450 (CYP), Loss-of-function (LoF) impair between individuals races. Therefore, testing effective strategy personalized incidence CAD patients. Purpose We sought develop a nested allele-specific multiplex polymerase chain reaction (PCR) method detection CYP2C9 LOF alleles. Method Genomic DNA was extracted from taken 7 healthy volunteers using commercial extraction kit. A two-step PCR developed. First, subjected first amplify exons 4 5 simultaneously single tube. Second, The products 1st were then as template second detect alleles (CYP2C19*2 *3) primers. test results validated via sequencing. Result successfully detected clinically (Figure 1). This fast doesn't require high-quality expensive laboratory equipment. It reproducible specific can reliably variants. Direct sequencing target variants been validate further, amplified sequences 100% identical sequence. Conclusion developed methods identify (rs4244285 rs4986893). understand ethnic differences their potential implications therapy, we highly recommend identifying relevant before clinical manifestations arise. Funding Acknowledgement Type funding sources: None.