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Rapid detection of tobacco mosaic virus using the reverse transcription loop-mediated isothermal amplification method

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Abstract

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 Mg2+, 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.

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Acknowledgments

This work was supported by the establishment of the tobacco germplasm platform of China and detection of important viruses of tobacco by LAMP from Chinese State Tobacco Monopoly Administration, People’s Republic of China. We thank Wang Fenglong and Dr. Yang Jinguang for providing virus samples and experimental help.

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Authors and Affiliations

  1. Qingzhou Tobacco Research Institute of CNTC, 11, 4th Road Keyuan Longitude, Laoshan District, Qingdao, Shandong, 266101, China

    Yanhua Liu, Zhide Wang, Yumei Qian, Jianmin Mu, Lili Shen, Fenglong Wang & Jinguang Yang

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  1. Yanhua Liu
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  2. Zhide Wang
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  3. Yumei Qian
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  4. Jianmin Mu
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  5. Lili Shen
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  6. Fenglong Wang
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  7. Jinguang Yang
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Corresponding author

Correspondence to Zhide Wang.

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Liu, Y., Wang, Z., Qian, Y. et al. Rapid detection of tobacco mosaic virus using the reverse transcription loop-mediated isothermal amplification method. Arch Virol 155, 1681–1685 (2010). https://doi.org/10.1007/s00705-010-0746-5

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  • DOI: https://doi.org/10.1007/s00705-010-0746-5

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