番茄不孕病毒实时荧光RT-PCR检测方法研究  

Detection of tomato aspermy virus by TaqMan real-time RT-PCR

作　　者：谭钟[1] 闻伟刚[1] 张吉红[1] 张颖[1] 

机构地区：[1]宁波出入境检验检疫局,宁波315012

出　　处：《安徽农业大学学报》2009年第4期655-658,共4页Journal of Anhui Agricultural University

基　　金：宁波出入境检验检疫局科技项目(甬K01-2008)资助

摘　　要：番茄不孕病毒(tomato aspermy virus,TAV)是危害我国菊花生产的主要病毒之一。本研究根据该病毒外壳蛋白北京分离物的基因,设计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DAS-ELIAS相比,检测灵敏度提高了约1 000倍,重复性试验表明批内与批间变异系数均在5%以内,整个检测过程仅需2 h,是一种操作简便、特异性强、灵敏度高、快速有效的TAV检测方法。Tomato aspermy virus （TAV） is one of the main chrysanthemum virus in China. In this study, a pair of primers and a probe were designed and synthesized based on the nucleotide sequence of coat protein gene from Beijing isolate. The result suggests that real-time RT-PCR assay showed about 1 000-fold sensitivity compared to that by DAS-ELIAS. The varying coefficiencies of intra-assay and inter-assay for quantitative detection were less than 5 %. The overall test could be completed within about 2 h. This system offers an easily, specific, sensitive, high throughput and rapid method for TAV detection.

关 键 词：番茄不孕病毒 实时荧光RT-PCR 检测 

分 类 号：S41-30[农业科学—植物保护]