多重荧光定量 PCR 检测麻疹和风疹病毒的效果评价  

Effect evaluation of detection of measles and rubella virus by multiplex fluorescent quantitative PCR

作　　者：房学强 许青[2] FANG Xue-qiang;XU Qing(Research and Development Center for Maternal and Child Health,Maternal and Child Health Care Hospital of Shandong Province,Jinan Shandong,250014,China;EPI Institute,Shandong Center for Disease Control and Prevention,Jinan Shandong,250014,China)

机构地区：[1]山东省妇幼保健院妇幼健康研究发展中心,山东济南250014 [2]山东省疾病预防控制中心免疫所,山东济南250014

出　　处：《职业与健康》2020年第21期2916-2919,2926,共5页Occupation and Health

基　　金：山东省医药卫生计划项目(2017WS453);山东省预防医学会智飞疾病预防控制技术研究基金(LYH2017-06)。

摘　　要：目的评价多重荧光定量聚合酶链式反应(PCR)检测麻疹和风疹病毒核酸的效果。方法对包括麻疹、风疹、水痘、登革热、流行性腮腺炎、艾滋病毒(HIV)、流感、乙肝、丙肝、乙脑10种病毒和阴性对照共143份标本分别提取核酸,进行多重荧光定量PCR试验和普通反转录PCR(RT-PCR)扩增麻疹、风疹病毒特异性片段并测序。对两种方法的检测结果进行比对,进行配对卡方McNemer检验和Kappa一致性检验。结果该多重荧光定量PCR方法的灵敏度为100.00%,特异度为94.67%,假阳性率为5.33%,假阴性率为0。多重荧光定量PCR和普通RT-PCR法一致性检验参数κ=0.94。结论该多重荧光定量PCR方法灵敏度高,特异性好,与其他常见病毒无交叉反应,且含有内标作为采样环节的质控,适于麻疹和风疹病毒的快速诊断。Objective To evaluate the efficiency of a multiplex fluorescent quantitative PCR method for the detection of measles and rubella virus nucleic acids.Methods The nucleic acids were extracted from 143 samples which included measles,rubella,varicella,dengue fever,mumps,HIV,influenza,hepatitis B,hepatitis C,JE and negative control.RT-PCR amplification,sequencing,genotyping of the specific fragments of measles and rubella virus and the multiplex fluorescent quantitative PCR were carried out.The results of the two methods were compared and matched by McNemer chi-square test and Kappa consistency test.Results The sensitivity,specificity,false-positive rate and false negative rate of the method were 100.00%,94.67%,5.33%and 0,respectively.The consistency test parameterκof the results of fluorescent quantitative PCR and RT-PCR was 0.94.Conclusion The multiplex fluorescent quantitative PCR method has high sensitivity and specificity,has no cross reaction with other common viruses,and contains internal standard as the quality control of sampling.It is suitable for rapid diagnosis of measles virus and rubella virus.

关 键 词：多重荧光定量PCR 麻疹病毒 风疹病毒 反转录PCR 基因测序 

分 类 号：R117[医药卫生—公共卫生与预防医学]