实时荧光RT-PCR方法检测齿兰环斑病毒与建兰花叶病毒  

Detection of Odontoglossum ringspot virus and Cymbidium mosaic virus by real-time fluorescent RT-PCR

作　　者：闻伟刚[1] 谭钟[1] 崔俊霞[1] 

机构地区：[1]宁波出入境检验检疫局,宁波315012

出　　处：《植物保护》2009年第4期101-104,共4页Plant Protection

基　　金：宁波出入境检验检疫局科技项目(甬K04-2005)

摘　　要：齿兰环斑病毒(Odontoglossum ringspot virus,ORSV)与建兰花叶病毒(Cymbidium mosaic virus,CymMV)是严重危害兰科植物的2种主要病毒。本研究根据病毒不同分离株外壳蛋白基因的保守序列,分别设计了ORSV与CymMV各自的特异性引物与荧光探针,建立了基于TaqMan探针的实时荧光RT-PCR检测方法。该方法与酶联免疫检测方法相比,灵敏度提高104倍,具有快速、灵敏和高特异性的优点,适合对ORSV和CymMV的快速检测。应用该方法,从来自台湾的蝴蝶兰样品中检出齿兰环斑病毒。Odontoglossum ringspot virus （ORSV） and Cymbidium mosaic virus （CymMV） are the two major types of viruses infecting the Orehidaceae severely. In this study, based on the conserved sequences in coat protein genes of ORSV and CymMV, the real-time fluorescent RT-PCR primers and probes specific to ORSV and CymMV were designed, and the real-time fluorescent RT-PCR methods were established. Compared to ELISA method, the real-time RT-PCR assay showed 10 000-fold higher than ELISA in detection sensitivity. It is a rapid, sensitive and highly specific method for detection of ORSV and CymMV. With this method, ORSV in phalaenopsis imported from Taiwan was successfully intercepted.

关 键 词：齿兰环斑病毒 建兰花叶病毒 实时荧光RT-PCR 检测 

分 类 号：S432.41[农业科学—植物病理学]