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

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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Correspondence to Jeremy R. Thompson.

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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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