半巢式RT-Realtime PCR检测南芥菜花叶病毒  

Dectection of Arabis Mosaic Virus by Semi-Nested RT-Realtime PCR

作　　者：粟智平 杨益娥 邓丛良[3] 李金庆 王颖 段效辉 

机构地区：[1]烟台出入境检验检疫局,山东烟台264000 [2]湖南省会同县农业局 [3]北京出入境检验检疫局

出　　处：《现代农业科技》2015年第18期145-147,共3页Modern Agricultural Science and Technology

基　　金：国家973研课题(2011CB932800);烟台市科学技术发展计划项目(2008324);山东检验检疫局科研项目(SK201305);山东检验检疫局科研项目(SK2011)

摘　　要：根据南芥菜花叶病毒(Ar MV)外壳蛋白(CP)基因的保守序列,设计合成了3条巢式PCR引物和1条Taq MAN荧光探针,建立了半巢式RT-Realtime PCR检测Ar MV的新方法。该方法有机地结合了巢式PCR和Taq MAN探针检测技术。第二步半巢式-RT-Realtime PCR既是对第一步信号的进一步放大,也是对第一步PCR产物的确认,因此,检测的准确性、灵敏度比巢式PCR、Realtime PCR等方法高。结果表明:该方法检测灵敏度可达0.06 fg/μL植物总RNA。Three nested primers and one TaqMAN probe were designed according to the CP gene of the Arabis mosaic virus ,with which we set up a new method of semi-nested RT-Realtime PCR to detect the ArMV.The nest-PCR and the probe-detection technique were combined maneuverably in this study.The semi-nested RT-Realtime PCR in the second step could both magnify the sigal and validate the result of the first step,which could provide a more sensitive and specific detection of the ArMV.The sensitivity of the method could reach 0.06 fg/μL of total plant RNA.

关 键 词：南芥菜花叶病毒(ArMV) 半巢式RT—ReMtimePCR 检测 

分 类 号：S41-30[农业科学—植物保护]