目的?对人细胞色素P450酶等位基因(CYP2C19)多态性检测试剂盒进行性能验证。方法?根据中国合格评定国家认可委员会CNAS-GL039:2019《分子诊断检验程序性能验证指南》相关要求，结合试剂盒说明书，从符合率、精密度、检出限和抗干扰能力4个方面对“人CYP2C19基因多态性检测试剂盒(荧光PCR法)”进行性能验证。符合率验证采用荧光PCR法和Sanger测序法检测；精密度验证选择杂合突变CYP2C19?c.681GA、c.636GA样本进行检测；抗干扰能力验证判断基因型检测是否受影响。结果?PCR法和Sanger测序法的检测符合率为100%；?CYP2C19?c.681 G和A等位基因的FAM和ROX荧光通道精密度检测CV值分别为3.78%、1.76%、3.76%和3.56%；CYP2C19?c.636 G和A等位基因的FAM和ROX荧光通道精密度检测CV值分别为3.08%、2.81%、3.40%和3.73%；样本最低检出限为10 ng/μL；抗干扰能力验证显示基因型可以正常检测。结论?人CYP2C19基因多态性检测试剂盒(荧光PCR法)符合ISO15189质量管理要求，可以为临床提供准确可靠的检测结果。
Objective?To verify the performance of the detection kits for human cytochrome P450 enzyme allele (CYP2C19) polymorphism.?Methods?According to China′s Conformity Assessment (CNAS) National Approval Committee CNAS-GL039: 2019 “Guidance on the Performance Verification for Molecular Diagnostic Procedures” combined with the reagent instruction, four aspects of performances for the human?CYP2C19?gene polymorphism detection kit (real-time quantitative PCR), i.e., coincidence rate, precision, detection limit and anti-interference ability, were verified. The coincidence rate was verified by fluorescence PCR and Sanger sequencing. The samples of heterozygous mutant?CYP2C19?c.681GA and c.636GA were selected to verify the precision. The anti-interference ability test was used to assess the interferences of genotype detection.?Results?The coincidence rate of the results of PCR and Sanger sequencing was 100%.?CV?values of precision verification of FAM and ROX fluorescence channels for?CYP2C19?c.681 alleles G and A were 3.78%, 1.76%, 3.76% and 3.56%, respectively.?CV?values of precision verification for FAM and ROX fluorescence channels of?CYP2C19?c.636 alleles G and A were 3.08%, 2.81%, 3.40% and 3.73%, respectively. The minimum detection limit of samples was 10 ng/μL. The verification for anti-interference ability showed that all the genotypes could be detected normally.?Conclusion?The kit for human?CYP2C19?gene polymorphism detection (real-time quantitative fluorescent PCR) conformed to ISO15189 quality management requirements and could provide accurate and reliable detection results for clinical practice.