Qingzhou Tobacco Research Institute of CNTC, 4th Road Keyuan Longitude, Laoshan District, Qingdao, Shandong, China.
Arch Virol. 2010 Oct;155(10):1681-5. doi: 10.1007/s00705-010-0746-5. Epub 2010 Aug 1.
Tobacco mosaic virus (TMV) can cause a severe disease that is capable of greatly reducing tobacco quality and yield. In this study, a one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detection of TMV. The concentration of Mg(2+), reaction temperature and reaction time of the RT-LAMP were optimized to 5 mM, 65°C, and 60 min, respectively. The detection limit of the method was 100 times higher than that of RT-PCR. Visual inspection of RT-LAMP amplification demonstrated that positive and negative reactions exhibit distinctly different colours in daylight. Our results demonstrate that the method is stable, sensitive and specific.
烟草花叶病毒（TMV）可引起严重疾病，极大地降低烟草的质量和产量。本研究建立了一种一步法逆转录环介导等温扩增（RT-LAMP）检测方法。优化了 Mg(2+)浓度、反应温度和反应时间，分别为 5 mM、65°C 和 60 min。该方法的检测限比 RT-PCR 高 100 倍。RT-LAMP 扩增的目视检查表明，阳性和阴性反应在日光下呈现明显不同的颜色。我们的结果表明该方法稳定、敏感且特异。
