Establishment Of CYP2C19 Gene Polymorphism Detection And Its Clinical Applications

Objective:Establish the CYP2C19 polymorphism detection method and to evaluate whether it can be used for clinical testing.Methods:Establish the CYP2C19 polymorphism detection method. According to the requirements of National Academy of Clinical Biochemistry (NACB) to develop the laboratory developed test-C7P2C19 genotypes. CYP2C19 genotypes were detected by PCR-fluorescent capillary electrophoresis sequencing technology. The limit of detection, accuracy, precision, analytical specificity and other parameters of the assay were validated. The Kappa test was used to evaluated accuracy. Coincidence rate was used to evaluate the repeatability.Sequencing quality verification based on fluorescent units (RFU), noise ratio, quality values (QV).From October 2014 to January 2015,120 patients with CAD who undergoing Percutaneous coronary intervention (PCI) in The First Affiliated Hospital of Kunming Medical University were enrolled in our study. According to genotype, patient were classified as:Extensive metabolizer(EM), Intermediate metabolizer(IM), and Poor metabolizer(PM). All patients were followed-up 12 month after PCI. Primary clinical endpoint was defined as major adverse cardiovascular events(MACE) and the secondary clinical endpoints were acute coronary syndrome(ACS). Compare the difference of clinical information in different metabolism group. Logistic regression analysis was used to determine the correction between MACE/ACS and possible risk factors.Results:1.when Ct≤32, the signal intensity was beyond 200 RFU, background peaks was not exceed 5% in a sequence; 20 times was performed repeatedly and all the samples can be detected. There was a close agreement between two laboratory(Kappa=1.00). The coincidence rate was 100%.2.The genotype distribution of CYP2C19*2:there were 57 patients carrying GG,52 patients carrying GA and 11 patients carrying AA. The genotype distribution of CYP2C19*3:there were 113 patients carrying GG,4 patients carrying GA and 3 patients carrying AA.3.There was no difference in clinical information.11 patients had MACE and 32 patients had ACS during 12-month follow-up.The significant difference appeared on the incidence of ACS and MACE in separate metabolism groups(P<0.05).4.Logistic aggression analysis shows patients with Diabetes had a high incidence of MACE compare with non-Diabetic patients(OR=0.163,95%CI 0.035-0.766, P=0.022); smokers had a low incidence of ACS campare with non-smokers(OR=5.273,95%CI 1.339-20.758; P=0.017); patients in IM and PM group had a high incidence both of MACE(OR=5.119,95%CI 1.276-20.541, P=0.021)and ACS(OR=4.779,95%CI 1.918-11.912; P=0.001) compare with patients in EM.Conclusions:CYP2C19 polymorphism detection method in this study can be used in clinical detection..