腮腺炎病毒临床口漱液样本的一步法实时荧光定量PCR检测

目的建立腮腺炎病毒(mumps virus,MuV)临床口漱液样本一步法荧光定量PCR检测方法,为腮腺炎的临床诊断及免疫防控提供依据。方法对101份采集自中国南部3个省、自治区(云南、四川、广西)的临床疑似腮腺炎患者口漱液样本,分别采用细胞培养-MuV特异的巢式PCR法及TaqMan探针Real-time PCR法进行检测,比较两种方法检测MuV的灵敏性。结果细胞培养-MuV特异的巢式PCR法共检出31份MuV阳性样本,系统进化分析表明,该31株MuV均属于F基因亚型;TaqMan探针Real-time PCR法共检出41份MuV阳性样本,其中包含了细胞培养-MuV特异的巢式PCR法检出的31份阳性样本,TaqMan探针Real-time RT-PCR法对MuV的检出率(40.59%)高于传统细胞培养-巢式PCR法的检出率(30.69%)。结论 TaqMan探针Real-time PCR法的灵敏性、特异性和简便性均优于传统的细胞培养-MuV特异的巢式PCR法,可应用于腮腺炎的临床诊断及免疫防控。

One-step real-time fluorescent quantitative PCR assay for mumps virus in clinical saliva specimens

Objective To develop an one-step real-time fluorescent quantitative PCR assay for mumps virus(MuV)in clinical saliva specimens and provide a basis for clinical diagnosis,prevention and control of mumps.Methods A total of 101 clinical saliva specimens collected from suspected patients with mumps in Yunnan and Sichuan Province as well as Guangxi Zhuang Autonomous Region,China were determined by cell culture-MuV specific nested PCR and real-time PCR with TaqMan probe respectively,based on which the sensitivities of the two methods were compared.Results Thirty-one MuV-positive specimens were found by cell culture-MuV specific nested PCR,all of which belonged to genotype F as proved by phylogenetic tree.However,41 MuV-positive specimens were found by real-time PCR with TaqMan probe,including the 31 positive specimens proved by cell culture-MuV specific nested PCR,indicating that the positive rate of Muv by real-time RT-PCR with TaqMan probe(40.59%)was higher than that by traditional cell culture-nested PCR(30.69%).Conclusion The sensitivity,specificity and simplicity of real-time PCR with TaqMan probe were higher than those of traditional cell culture-MuV specific nested PCR,which might be used for the clinical diagnosis,prevention and control of mumps.