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Reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid diagnosis of chilli veinal mottle virus

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Abstract

Chilli veinal mottle virus (ChiVMV) causes significant economic loss to chilli cultivation in northeastern India, as well as in eastern Asia. In this study, we have developed a single-tube one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid, sensitive and specific diagnosis of ChiVMV. Amplification could be visualized after adding SYBR Green I (1000×) dye within 60 min under isothermal conditions at 63 °C, with a set of four primers designed based on the large nuclear inclusion protein (NIb) domain of ChiVMV (isolate KC-ML1). The RT-LAMP method was 100 times more sensitive than one-step reverse transcription polymerase chain reaction (RT-PCR), with a detection limit of 0.0001 ng of total RNA per reaction.

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Acknowledgment

The work was supported by IXX08592 project of the Indian Council of Agricultural Research (ICAR). We thank Prof. F. Murilo Zerbini for critically reviewing this manuscript, and the anonymous reviewers for their comments and suggestions.

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Authors and Affiliations

  1. ICAR Research Complex for North Eastern Hill Region, Umiam, Meghalaya, 793 103, India

    Amrita Banerjee, Satish Chandra & SV Ngachan

  2. ICAR-National Bureau of Plant Genetic Resources, Regional Station, Umiam, Meghalaya, 793 103, India

    Somnath Roy

  3. ICAR Research Complex for NEH Region, Manipur Centre, Lamphelpat, Manipur, 795 004, India

    Susheel Kumar Sharma

  4. ICAR Research Complex for NEH Region, Mizoram Centre, Kolasib, Mizoram, 796 081, India

    Sudip Kumar Dutta

Authors
  1. Amrita Banerjee
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  2. Somnath Roy
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  3. Susheel Kumar Sharma
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  4. Sudip Kumar Dutta
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  5. Satish Chandra
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  6. SV Ngachan
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Correspondence to Amrita Banerjee.

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Banerjee, A., Roy, S., Sharma, S.K. et al. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid diagnosis of chilli veinal mottle virus. Arch Virol 161, 1957–1961 (2016). https://doi.org/10.1007/s00705-016-2850-7

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  • DOI: https://doi.org/10.1007/s00705-016-2850-7

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