荧光PCR检测高血压用药相关基因多态性方法的建立  

Establishment of real-time PCR assay for detection of hypertension drug-related gene polymorphism

作　　者：刘秀卿[1] 李延武[1] 黄磊[1] LIU Xiuqing;LI Yanwu;HUANG Lei(Department of Laboratory Medicine,The Second People’s Hospital of ShenZhen,ShenZhen,GuangDong,China,518035)

机构地区：[1]深圳市第二人民医院检验科,广东深圳518035

出　　处：《分子诊断与治疗杂志》2021年第5期707-711,共5页Journal of Molecular Diagnostics and Therapy

摘　　要：目的建立联合检测高血压用药相关基因多态性的荧光PCR检测方法。方法针对CYP2D6*10、CYP2C9*3、ADRB1(1165G>C)、AGTR1(1166A>C)、ACE(I/D)突变位点分别设计特异性引物和探针,优化建立荧光PCR反应体系。验证该方法的敏感性、特异性、准确度、检出限、抗干扰能力。收集收集2017年6月至2019年1月本院EDTA-K2抗凝全血样本362例临床样本,分别用荧光PCR法、基因芯片法、测序法进行检测,以基因芯片法为对比方法,测序法为金标准,评价荧光PCR法的特异性和准确性。结果建立的荧光PCR法对五个位点的检测CV值均小于5%,检出限分别为5、10、20、5、20 ng/m L。362例临床样本的检测结果与测序法的一致性为100%,敏感性、特异性、准确性均高于基因芯片法。结论建立的联合检测高血压用药基因多态性的荧光PCR法具有很好的准确性、特异性、灵敏度和重复性,且操作简单、结果判读容易,可为高血压药物的临床使用提供参考。Objective To establish a real-time PCR assay for detecting drug-related genes polymorphism in hypertension.Methods A set of allele-specific PCR primers and probes were designed for CYP2 D6*10,CYP2 C9*3,ADRB1(1165 G>C),AGTR1(1166 A>C),ACE(I/D).A real-time PCR reaction system was established and optimized.The accuracy,susceptibility,specificity and repeatability of the assay was evaluated.A total of 362 clinical samples of EDTA-K2 anticoagulant whole blood samples from Shenzhen Second People.s Hospital from June 2017 to January 2019 were collected,and tested with fluorescence PCR,gene chip method,and sequencing method respectively.The gene chip method was used as the comparison method,and the sequencing method was used as the gold standard to evaluate the specificity and accuracy of the fluorescent PCR method.Results The established fluorescence PCR method has detected CV values of less than 5%for the five loci,and the detection limits are 5、10、20、5、20 ng/m L.The consistency of the test results of 362 clinical samples with the sequencing method was 100%,and the sensitivity,specificity,and accuracy were higher than those of the gene chip method.Conclusion The established fluorescent PCR method for detecting gene polymorphisms of hypertension drugs has good accuracy,specificity,sensitivity and reproducibility,simple operation and easy interpretation of results,which can provide reference for the clinical use of hypertension drugs.

关 键 词：高血压 基因多态性 CYP2D6 CYP2C9 ADRB1 AGTR1 ACE 

分 类 号：R544.1[医药卫生—心血管疾病]