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实时荧光PCR检测西尼罗病毒方法的建立

作  者:
邱璐;李小林;蒋静;王艳;周小俊
单  位:
上海出入境检验检疫局;中国疾病预防控制中心寄生虫病预防控制所
关键词:
西尼罗病毒;实时荧光RT-PCR;特异性;敏感性
摘  要:
西尼罗病毒可引起人畜共患虫媒病,已经在全球许多国家流行。主要随迁飞的鸟类携带扩散,传入我国的风险很大。本研究在参考多株西尼罗病毒基因序列的基础上,设计引物及探针,建立了实时荧光RT-PCR的方法。特异性试验表明,该方法对同为黄病毒科的日本脑炎病毒(JEV)SA14-14-2株和登革病毒(DEN)Ⅰ型cDNA检测均为阴性;敏感性试验证实,实时RT-PCR检测敏感度要比常规RT-PCR法高100倍。说明本研究建立的实时RT-PCR方法敏感性和特异性均较高,可用于西尼罗病毒感染的早期监测和诊断。
译  名:
Detection of West Nile virus by real-time RT-PCR
作  者:
QIU Lu1,LI Xiao-lin1,JIANG Jing1,WANG Yan1,ZHOU Xiao-jun2(1.Shanghai Entry-Exit Inspection and Quarantine Bureau,Shanghai 200135,China; 2.National Institute of Parasitic Diseases,Chinese Center for Disease Control and Prevention, Shanghai 200025,China)
关键词:
West Nile virus;real-time RT-PCR;specificity;sensitivity
摘  要:
West Nile virus(WNV) is by far the most widely distributed arbovirus.The WNV infection has been recognized as an emerging infectious disease with possible transmitting risk in our country.The pairs of primers and TaqMan probes were designed and synthesized based on the nucleotide sequences of WNV in GenBank.After optimization,the real-time PCR was established.These primers were applied to detect the nucleic acids of WNV,Japanese B encephalitis virus(JEV) and Dengue viruses(DEN),and the latter two viruses could not be detected.Compared with conventional RT-PCR,real-time PCR appeared to be approximately 100 times more sensitive in detection of WNV.It suggests that the real-time PCR developed for the detection of WNV shows high degree of specificity and sensitivity,and can be used for WNV inspection and investigation.

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