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闻伟刚1,杨翠云2,崔俊霞1,张颖1.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.
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
  
DOI:
中文关键词:  
英文关键词:Bean pod mottle virus (BPMV)  loop mediated isothermal amplification (LAMP)  detection
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作者单位
闻伟刚1,杨翠云2,崔俊霞1,张颖1  
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中文摘要:
      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。
英文摘要:
      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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