豇豆重花叶病毒RT-LAMP检测方法的建立

摘要

豇豆重花叶病毒(Cowpea severe mosaic virus,CPSMV)是我国进境植物检疫性有害生物,本研究根据CPSMV的外壳蛋白基因序列设计引物,以期建立CPSMV的反转录-环介导等温扩增(reverse transcription loopmediated isothermal amplification,RT-LAMP)检测方法.条件优化后,建立了CPSMV的RT-LAMP方法,该方法可将CPSMV与同属其他病毒准确区分,具有良好的特异性.灵敏度试验显示,RT-LAMP比普通RT-PCR灵敏度高10倍.说明本研究建立的RT-LAMP方法适用于CPSMV的快速、特异和灵敏的检测.%Cowpea severe mosaic virus(CPSMV)is a quarantine pathogen in China. This study is aimed to establish a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for detection of CPSMV. The results showed that, under optimal reaction conditions, the RT-LAMP method could distinguish CPSMV from other comoviruses. A sensitivity test showed that this method was 10 times more sensitive than the standard RT-PCR assay. The method in this study is suitable for fast, specific and sensitive detection of CPSMV.