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Rapid detection of tobacco viruses by reverse transcription loop-mediated isothermal amplification

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Abstract

Tobacco viruses may cause a wide range of diseases that heavily reduce tobacco quality and yield worldwide. In order to detect viral diseases in tobacco fields, a one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) method was established. Nucleotide amplification could be observed clearly after adding SYBR Green I, within 60 min under isothermal conditions, at 63-65 °C with a set of primers targeting the viral coat protein (CP) genes of tobacco viruses including cucumber mosaic virus (CMV), potato virus Y (PVY), tobacco etch virus (TEV), tobacco mosaic virus (TMV) and tobacco vein banding mosaic virus (TVBMV). This method has high specificity and sensitivity. The sensitivity of the RT-LAMP was 10 to 100 times higher than that of the conventional RT-PCR method. The RT-LAMP assay was proven reliable for virus diagnosis of tobacco samples from the field.

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Acknowledgments

The authors would like to thank Wenjun Zhao (of Chinese Academy of Inspection and Quarantine, Beijing, China), Zhanmin Wu, Jin Dai and Ting Huang (of State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A&F University) for kindly providing the tobacco virus samples. This study was supported by 111 Project of Education Ministry of China (B07049) and Key Program for Science and Technology Development of China National Tobacco Corporation (110200902046).

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Authors and Affiliations

  1. Key Laboratory of Crop Pest Integrated Pest Management on Crop in Northwestern Loess Plateau, Ministry of Agriculture, Key Laboratory of Plant Protection Resources and Pest Management, Ministry of Education, State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A&F University, Yangling, 712100, People’s Republic of China

    Lei Zhao, Xingan Hao & Yunfeng Wu

  2. Shaanxi Tobacco Research Institute, Xi’an, 710003, Shaanxi, China

    Julong Cheng

  3. Yangling Vocational and Technical College, Yangling, 712100, Shaanxi, China

    Xiaoman Tian

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  1. Lei Zhao
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  2. Julong Cheng
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  3. Xingan Hao
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  4. Xiaoman Tian
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  5. Yunfeng Wu
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Corresponding author

Correspondence to Yunfeng Wu.

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Zhao, L., Cheng, J., Hao, X. et al. Rapid detection of tobacco viruses by reverse transcription loop-mediated isothermal amplification. Arch Virol 157, 2291–2298 (2012). https://doi.org/10.1007/s00705-012-1441-5

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  • DOI: https://doi.org/10.1007/s00705-012-1441-5

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