逆转录环介导等温扩增技术检测烟草脆裂病毒

Detection of tobacco rattle virus by reverse-transcription loop-mediated isothermal amplification

为实现进境种球中烟草脆裂病毒(Tobacco rattle virus,TRV)的快速检测,本研究应用逆转录环介导等温扩增技术,建立了TRV-RT-LAMP检测方法,在60℃,60 min条件下,能特异性地检测到TRV,灵敏度可达到10 pg/μL,是普通RT-PCR的100倍。用SYBR Green I染色后可以使检测结果可视化,方便快速。同时结合RNA粗提,建立了一步法RT-LAMP技术,可以快速检测植株中的TRV。

In order to detect tobacco rattle virus(TRV)from imported bulbs rapidly,this study establishes a TRV-RT-LAMP detection method by using reverse transcription loop-mediated isothermal amplification technique.TRV can be specifically detected at 60℃in 60 min,with a sensitivity of 10 pg/μL,which is 100 times of ordinary RT-PCR.After dyeing with SYBR Green I,the detection results can be visualized.A one-step RT-LAMP method is also established to detect TRV in plants combining with RNA crude extraction.