基于TaqMan MGB探针的花生矮化病毒检测研究  

Detection of Peanut stunt virus using TaqMan MGB probe

作　　者：闻伟刚[1] 谭钟[1] 张颖[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》2010年第3期121-124,共4页Plant Protection

基　　金：宁波出入境检验检疫局科技项目(甬K01-2009)

摘　　要：花生矮化病毒(Peanut stunt virus,PSV)是我国进境检疫性有害生物。本研究根据该病毒不同分离株外壳蛋白基因(coat protein,CP)的保守序列,设计了特异性引物与TaqMan MGB荧光探针,建立了PSV的实时荧光RT-PCR检测方法。方法特异性研究表明,针对PSV的2个不同株系PSV-E和PSV-W,均能够得到典型扩增曲线,Ct值分别为20.10和21.22;而对于黄瓜花叶病毒、番茄不孕病毒、马铃薯Y病毒、菜豆荚斑驳病毒以及烟草环斑病毒等其他毒株则没有典型扩增曲线,也无Ct值。灵敏度比较发现,该方法比普通RT-PCR检测方法的灵敏度提高100倍,具有快速、灵敏和高特异性的优点,适合对PSV的检测。Peanut stunt virus （PSV） has been listed as an important quarantine pest by Chinese government. In this study, a real-time fluorescent RT-PCR method was established based on specific primers and TaqMan MGB probe designed in accordance with the conserved sequences of the coat protein gene of different PSV isolates. Specificity studies showed that for the two different PSV strains PSV-E and PSV-W, a typical amplification curve can be obtained, with Ct values as 20.10 and 21.22, respectively. No typical amplification curves and Ct values could be obtained for the Cucumber mosaic virus, Tomato aspermy virus, Potato virus Y, Bean pod mottle virus, Tobacco ringspot virus and other virus strains. Sensitivity comparisons indicated that the method could improve sensitivity by 100 times compared with regular RT-PCR method. The real-time fluorescent RT-PCR was a rapid, sensitive and highly specific method for detection of PSV.

关 键 词：花生矮化病毒 TAQMAN MGB探针 实时荧光RT-PCR 检测 

分 类 号：S432.41[农业科学—植物病理学]