CYP2C19基因多态性荧光定量PCR检测方法的建立  

Establishment of real-time PCR assay for the detection of CYP2C19 gene polymorphism

作　　者：刘秀卿[1] 李卓成[1] 李延武[1] 

机构地区：[1]深圳市第二人民医院检验科,广东深圳518035

出　　处：《临床检验杂志》2016年第2期95-99,共5页Chinese Journal of Clinical Laboratory Science

摘　　要：目的建立检测人细胞色素氧化酶2C19(CYP2C19)基因多态性的荧光定量PCR检测方法。方法用Primer Premier5.0和Primer Express 3.0软件针对人CYP2C19基因681和636多态性位点分别设计1对引物和2条Taq Man-MGB探针,优化建立荧光PCR反应体系。评价该方法的准确性、灵敏度和重复性。用该方法检测300例临床样本,并用测序法进行验证。结果建立CYP2C19基因681和636位点荧光PCR检测方法,灵敏度为每个反应2 ng/g DNA。300例临床样本检测结果与测序法一致性为100%。结论建立的CYP2C19基因多态性荧光PCR检测法准确性较高、灵敏度和重复性较好,适于临床实验研究。Objective To establish a real-time PCR method for the detection of cytochrome P450 2C19（ CYP2C19） gene polymorphism. Methods A pair of primers and two Taq Man-MGB probes for human CYP2C19 gene polymorphism loci 681 and 636 were designed using the Primer Premier 5. 0 and Primer Express 3. 0 software,and the established real-time PCR method was further optimized. Then,the accuracy,sensitivity and reproducibility of the established method were evaluated,300 clinical samples were detected by the method,and the results were further verified by the sequencing method. Results The sensitivity of the established real-time PCR assay for the detection of CYP2C19 polymorphisms loci 681 and 636 was about 2 ng / g DNA. The consistency of the results between the real-time PCR assay and the sequencing method was 100%. Conclusion The established real-time PCR assay for the detection of CYP2C19 gene polymorphim has good accuracy,sensitivity and reproducibility,which may be applied to the clinical research.

关 键 词：细胞色素P450 基因多态性 荧光聚合酶链式反应 检测方法 

分 类 号：R34[医药卫生—基础医学] R446