Rapid detection of tobacco mosaic virus using the reverse transcription loop-mediated isothermal amplification method.
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.
Liu
Yanhua
Y
Qingzhou Tobacco Research Institute of CNTC, 4th Road Keyuan Longitude, Laoshan District, Qingdao, Shandong, China.
Wang
Zhide
Z
Qian
Yumei
Y
Mu
Jianmin
J
Shen
Lili
L
Wang
Fenglong
F
Yang
Jinguang
J
D002413:Cations, Divalent / Q000378:metabolism; D003067:Coenzymes / Q000378:metabolism; D003116:Color; D008274:Magnesium / Q000378:metabolism; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010935:Plant Diseases / Q000821:virology; D048348:Reverse Transcription; D013696:Temperature; D013997:Time Factors; D014026:Nicotiana / Q000821:virology; D014027:Tobacco Mosaic Virus / Q000302:isolation & purification*; D014773:Virology / Q000379:methods*
D002413:Cations, Divalent; D003067:Coenzymes; D008274:Magnesium