Real-time RT-PCR检测西尼罗病毒一步法的建立  

Development of one step real-time RT-PCR method for detection of West Nile virus

作　　者：陈晓[1] 曾志磊[1] 朱丹[1] 彭丹[1] 余建萍[1] 谢鹏[1] 

机构地区：[1]重庆医科大学附属第一医院神经内科重庆医科大学神经科学研究中心,重庆400016

出　　处：《中国人兽共患病学报》2008年第12期1107-1110,共4页Chinese Journal of Zoonoses

基　　金："十一五"国家高科技发展计划(863计划)(No.2006AA02Z196)资助

摘　　要：目的建立针对西尼罗病毒E基因的一步法Real-time RT-PCR检测方法,用于西尼罗病毒感染样本的快速准确检测和定量。方法针对西尼罗病毒的保守基因E设计引物和探针,建立一步法Real-time RT-PCR反应方法并分析敏感性和特异性。结果本研究建立的一步法Real-time RT-PCR方法可以特异性检测出西尼罗病毒,不与日本脑炎病毒及其他病毒产生交叉反应。该方法的最低检出浓度为3.6×102copies/μl,标准曲线的线性范围为3.6×102～3.6×107copies/μl。结论本研究建立的一步法real-time RT-PCR方法敏感性和特异性较高,且不易出现污染引起的假阳性结果,适合用于西尼罗病毒感染样本的检测。One step real-time RT-PCR（Taqman） assays of West Nile virus（WNV） envelope protein gene was developed so that WNV RNA in field specimens or laboratory material can be characterized rapidly and specifically. A pair of specific oligonucleotide primers was designed. The linearity of the standard curve allowed quantification of 10^2 to 107 RNA transcripts/ul. and sensitivity of the test was close to 3.6 ×10^2 transcripts/ul. This assays were high specific without cross-reaction to Japanese encephalitis virus or other virus. This standardized technique is sensitive and reliable and allows rapid detection and quantiration of West Nile virus RNA in both field and experimental materials used for the surveillance and specific diagnosis of West Nile virus.

关 键 词：西尼罗病毒 real—time RT—PCR 一步法 TAQMAN探针 快速检测 

分 类 号：R373[医药卫生—病原生物学]