Article (Scientific journals)
New sensitive and fast detection of Little cherry virus 1 using loop-mediated isothermal amplification (RT-LAMP)
Tahzima, Rachid; Foucart, Y.; Peusens, G. et al.
2019In Journal of Virological Methods, 265, p. 91-98
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Keywords :
Closteroviridae; Diagnostics; LAMP; Little cherry virus; Plant virus; Velarivirus
Abstract :
[en] Little cherry virus 1 (LChV-1) belongs to the genus Velarivirus, family Closteroviridae, is an economically important pathogen affecting mainly cherry around the world emphasizing the impetus for its efficient and accurate on-site detection. This study describes the development of a reliable diagnostic protocol of LChV-1 based on a one-step reverse-transcription loop-mediated isothermal amplification (RT-LAMP). The protocol detects LChV-1 isolates in less than 10 min by fluorescence monitoring using a mobile detection device and is most optimal when performed at 67 °C. Sharp melting curves and unique melting temperatures (Tm) were obtained for the positive samples. Both the RT-LAMP and classical RT-PCR methods are capable of specifically detecting LChV-1 in infected leaf tissues. In addition, the RT-LAMP has remarkable advantages in comparison to RT-PCR. It is at least hundred fold more sensitive, significantly faster (allowing on-field leaf-to-result diagnostic) and efficient at minimal cost. In conclusion, this innovative RT-LAMP approach can contribute to the implementation of sustainable integrated management strategies for detection of LChV-1 in commercial orchards or for horticultural research stations. It is also suitable for decision support in phytosanitary epidemiological programs. © 2019 Elsevier B.V.
Disciplines :
Phytobiology (plant sciences, forestry, mycology...)
Author, co-author :
Tahzima, Rachid ;  Plant Sciences Unit, Flanders Research Institute for Agriculture, Fisheries and Food (ILVO), Merelbeke, 9820, Belgium, Laboratory of Integrated and Urban Phytopathology, University of Liège (ULg), Gembloux Agro-Bio tech, Gembloux, 5030, Belgium
Foucart, Y.;  Plant Sciences Unit, Flanders Research Institute for Agriculture, Fisheries and Food (ILVO), Merelbeke, 9820, Belgium
Peusens, G.;  Department of Zoology, Proefcentrum Fruitteelt (pcfruit), Sint-Truiden, 3800, Belgium
Beliën, T.;  Department of Zoology, Proefcentrum Fruitteelt (pcfruit), Sint-Truiden, 3800, Belgium
Massart, Sébastien  ;  Université de Liège - ULiège > Agronomie, Bio-ingénierie et Chimie (AgroBioChem) > Gestion durable des bio-agresseurs
De Jonghe, K.;  Plant Sciences Unit, Flanders Research Institute for Agriculture, Fisheries and Food (ILVO), Merelbeke, 9820, Belgium
Language :
English
Title :
New sensitive and fast detection of Little cherry virus 1 using loop-mediated isothermal amplification (RT-LAMP)
Publication date :
2019
Journal title :
Journal of Virological Methods
ISSN :
0166-0934
eISSN :
1879-0984
Publisher :
Elsevier B.V.
Volume :
265
Pages :
91-98
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 26 June 2019

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