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Arch Virol. 2019 May;164(5):1453-1457. doi: 10.1007/s00705-019-04207-y. Epub 2019 Mar 20.

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

Author information

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.
2
Plant Pathology and Plant-Microbe Biology Section, School of Integrative Plant Science, Cornell University, Ithaca, NY, 14853, USA.
3
Dept. de Genética Molecular y Microbiología, Pontificia Universidad Católica de Chile, Av. Libertador Bernardo O'Higgins 340, Santiago, Chile.
4
Plant Pathology and Plant-Microbe Biology Section, School of Integrative Plant Science, Cornell University, Ithaca, NY, 14853, USA. jrt36@cornell.edu.

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.

PMID:
30895404
DOI:
10.1007/s00705-019-04207-y

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