Polymorphism Detection of VKORC1 and CYP2C9 Genes for Warfarin Dose Adjustment by Real-Time PCR

Background: Managing individual-based anti-coagulant therapy with warfarin is an important goal for bleeding prevention in personalized medicine. Objectives: The purpose of the present study was to evaluate the allelic frequency of VKORC1 (C1173T, G1639A) and CYP2C9 (CYP2C9*2 and CYP2C9*3) polymorphic genes with SimpleProbeR real-time PCR as a fast, accurate and easy-to-handle method. Patients and Methods: One hundred unrelated patients under warfarin therapy were recruited as the study population. Real-time PCR was performed with SimpleProbeR for single-nucleotide polymorphisms (SNP) detection of CYP2C9 and VKORC1 genes. Results: The allelic frequencies for VKORC1 C1173T (CC, CT, TT) were 25%, 51% and 24%, respectively and for VKORC1 G1639A (-GG, GA, AA) were 27%, 51% and 22%, respectively. Despite a significant association between allelic polymorphisms in VKORC1 and warfarin dose (P 0.001), it was not statistically significant for CYP2C9*2 and CYP2C9*3, most probably due to the low rate of CYP2C9*2 and CYP2C9*3 SNPs observed. Conclusions: SimpleProbeR real-time PCR is a fast and accurate technique qualified to detect VKORC1 and CYP2C9 SNPs. These results encourage taking further steps towards using VKORC1 and CYP2C9 allelic screening to prevent clinical complications due to resistance or sensitivity to warfarin as well as anti-coagulant dose adjustment.