RT-LAMP技术检测菜豆荚斑驳病毒的研究  

Detection of Bean pod mottle virus by RT-LAMP

作　　者：闻伟刚[1] 杨翠云[2] 崔俊霞[1] 张颖[1] 

机构地区：[1]宁波出入境检验检疫局,宁波315012 [2]上海出入境检验检疫局,上海200135

出　　处：《植物保护》2010年第6期139-141,146,共4页Plant Protection

基　　金：宁波出入境检验检疫局科技项目(甬K01-2008)

摘　　要：菜豆荚斑驳病毒（Beanpodmottlevirus，BPMV）是我国对外公布的检疫性有害生物，本研究采用环介导等温扩增技术（100p-mediatedisothermal amplification，LAMP），建立了一种快速、灵敏和特异的BPMV检测方法。根据BPMV外壳蛋白编码基因上的8个位点，共设计了6条引物，通过RT-LAMP扩增得到特征性的瀑布状条带。特异性试验表明，引物对BPMV的检测具有良好的特异性；灵敏度试验显示RT-LAMP比RTPCR灵敏度高1000倍。该方法无需特殊的试剂和设备，只需在水浴锅中60℃等温扩增，整个检测周期约2～2．5h，适合BPMV的快速、准确检测。Bean pod mottle virus （BPMV） is a quarantine pest as approved by Chinese government. A rapid, sensitive and specific BPMV detection method using loop-mediated isothermal amplification （LAMP） was established in this study. Based on the sequence of BPMV coat protein coding gene, six primers were designed and the characteristic waterfall-like bands were obtained by RT-LAMP. Specificity tests showed that the primers were specific to BPMV; sensitivity test showed that RT-LAMP was 1000 times higher than RT-PCR. The method, without special reagents and equipments, was conducted with 60℃ isothermal amplification in a water bath and the entire testing cycle was about 2 2.5 h. It is suitable for rapid and accurate detection of BPMV.

关 键 词：菜豆荚斑驳病毒 环介导等温扩增 检测 

分 类 号：S432.41[农业科学—植物病理学] S436.43[农业科学—农业昆虫与害虫防治]