| RT-LAMP技术检测菜豆荚斑驳病毒的研究 |
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| 引用本文： | 闻伟刚,杨翠云,崔俊霞,张颖.RT-LAMP技术检测菜豆荚斑驳病毒的研究[J].植物保护,2010,36(6):139-141. |
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| 作者姓名： | 闻伟刚 杨翠云 崔俊霞 张颖 |
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| 作者单位： | 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 |
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| 基金项目： | 宁波出入境检验检疫局科技项目 |
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| 摘 要： | 菜豆荚斑驳病毒(Bean pod mottle virus,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(loop-mediated isothermal 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RT-PCR灵敏度高1 000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2~2.5 h,适合BPMV的快速、准确检测。
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| 关 键 词： | 菜豆荚斑驳病毒 环介导等温扩增 检测 |
Detection of Bean pod mottle virus by RT-LAMP |
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| Wen Weigang,Yang Cuiyun,Cui Junxia,Zhang Ying.Detection of Bean pod mottle virus by RT-LAMP[J].Plant Protection,2010,36(6):139-141. |
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| Authors: | Wen Weigang Yang Cuiyun Cui Junxia Zhang Ying |
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| Institution: | 1. Ningbo Entry Exit Inspection and Quarantine Bureau, Ningbo315012, China; 2. Shanghai Entry Exit Inspection and Quarantine Bureau, Shanghai200135, China |
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| Abstract: | 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. |
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| Keywords: | Bean pod mottle virus (BPMV) loop mediated isothermal amplification (LAMP) detection |
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