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J Virol Methods. 2014 Sep 1;205:24-30. doi: 10.1016/j.jviromet.2014.04.015. Epub 2014 May 4.

Rapid and sensitive detection of Little cherry virus 2 using isothermal reverse transcription-recombinase polymerase amplification.

Author information

1
Department of Plant Pathology, Washington State University, IAREC, 24106 North Bunn Road, Prosser, WA 99360, USA.
2
Agdia, Inc., 30380 County Road 6, Elkhart, IN 46514, USA.
3
Department of Plant Pathology, Washington State University, IAREC, 24106 North Bunn Road, Prosser, WA 99360, USA. Electronic address: keastwell@wsu.edu.

Abstract

Little cherry virus 2 (LChV2) (genus Ampelovirus) is the primary causal agent of little cherry disease (LCD) in sweet cherry (Prunus avium) in North America and other parts of the world. This mealybug-transmitted virus does not induce significant foliar symptoms in most sweet cherry cultivars, but does cause virus-infected trees to yield unevenly ripened small fruits with poor flavor. Most fruits from infected trees are unmarketable. In the present study, an isothermal reverse transcription-recombinase polymerase amplification (RT-RPA) technique was developed using LChV2 coat protein specific primers and probe. Detection of terminally labeled amplicons was achieved with a high affinity lateral flow strip. The RT-RPA is confirmed to be simple, fast, and specific. In comparison, although it retains the sensitivity of RT-PCR, it is a more cost-effective procedure. RT-RPA will be a very useful tool for detecting LChV2 from crude extracts in any growth stage of sweet cherry from field samples.

KEYWORDS:

Isothermal detection; Little cherry disease; Little cherry virus 2; Prunus avium; Recombinase polymerase amplification; Sweet cherry

PMID:
24797461
DOI:
10.1016/j.jviromet.2014.04.015
[Indexed for MEDLINE]
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