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A rapid, sensitive and inexpensive method for detection of grapevine red blotch virus without tissue extraction using loop-mediated isothermal amplification

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Abstract

Grapevine red blotch virus (GRBV) is an emerging virus of significant viticultural importance throughout North America. Here, we report the development of a simple protocol for point-of-use detection of GRBV. Extraction of nucleic acids is not required; instead, the whole intact plant can simply be pricked with a sterile pipette tip, which is then incubated in sterile distilled water to provide the sample template in a loop-mediated isothermal amplification (LAMP) reaction. This method is 10,000 times more sensitive than conventional PCR, costs under a dollar per sample, and can be completed from sampling to readout in just over half an hour.

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Acknowledgements

Many thanks to the Cornell Latin American Studies Program Fellowship for funding JLR2 while at Cornell. We are also grateful to John Keeton and colleagues at the USDA-ARS Plant Genetic Resources Unit for providing us with dormant field cane material from the USDA grapevine germplasm repository, and to Heather McLane, Aisha Younas and Alex Cha for their technical and greenhouse support. Thanks also to Marc Fuchs for critically reading the manuscript. This work was supported by USDA-NIFA Hatch project number 1013513, and the College of Agriculture and Life Sciences, Cornell University.

Funding

This study was funded by a Cornell Latin American Studies Program Fellowship, USDA-NIFA Hatch project number 1013513 and the College of Agriculture and Life Sciences, Cornell University.

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Authors and Affiliations

  1. Departamento de Biotecnología Agrícola, Instituto Politécnico Nacional, CIIDIR, Unidad Sinaloa, Blvd. Juan de Dios Bátiz Paredes No. 250, San Joachín, C.P. 81101, Guasave, Sinaloa, Mexico

    J. Lucina Romero Romero

  2. Plant Pathology and Plant-Microbe Biology Section, School of Integrative Plant Science, Cornell University, Ithaca, NY, 14853, USA

    Gavriela Dena Carver, Keith L. Perry & Jeremy R. Thompson

  3. Dept. de Genética Molecular y Microbiología, Pontificia Universidad Católica de Chile, Av. Libertador Bernardo O’Higgins 340, Santiago, Chile

    Patricio Arce Johnson

Authors
  1. J. Lucina Romero Romero
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  2. Gavriela Dena Carver
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  3. Patricio Arce Johnson
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  4. Keith L. Perry
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  5. Jeremy R. Thompson
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Corresponding author

Correspondence to Jeremy R. Thompson.

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All authors declare they have no conflict of interest.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Romero Romero, J.L., Carver, G.D., Arce Johnson, P. et al. A rapid, sensitive and inexpensive method for detection of grapevine red blotch virus without tissue extraction using loop-mediated isothermal amplification. Arch Virol 164, 1453–1457 (2019). https://doi.org/10.1007/s00705-019-04207-y

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  • DOI: https://doi.org/10.1007/s00705-019-04207-y

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