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Title:LAMP assay and rapid sample preparation method for on-site detection of flavescence dorée phytoplasma in grapevine
Authors:ID Kogovšek, Polona (Author)
ID Hodgetts, Jennifer (Author)
ID Hall, J. (Author)
ID Prezelj, Nina (Author)
ID Nikolić, Petra (Author)
ID Mehle, Nataša (Author)
ID Lenarčič, Rok (Author)
ID Rotter, Ana (Author)
ID Dickinson, M. (Author)
ID Boonham, Neil (Author)
ID Dermastia, Marina (Author)
ID Ravnikar, Maja (Author)
Files:URL URL - Source URL, visit http://onlinelibrary.wiley.com/doi/10.1111/ppa.12266/abstract;jsessionid=7850B082A5DCFD15057B22A9A431AFE1.f03t02
 
.pdf PDF - Presentation file, download (483,90 KB)
MD5: 658F0BDBF5F2A1BEA44A1431455F8AD8
 
URL URL - Source URL, visit https://doi.org/10.1111/ppa.12266
 
Language:English
Typology:1.01 - Original Scientific Article
Organization:Logo NIB - National Institute of Biology
Abstract:In Europe the most devastating phytoplasma associated with grapevine yellows (GY) diseases is a quarantine pest, flavescence dorée (FDp), from the 16SrV taxonomic group. The on-site detection of FDp with an affordable device would contribute to faster and more efficient decisions on the control measures for FDp. Therefore, a real-time isothermal LAMP assay for detection of FDp was validated according to the EPPO standards and MIQE guidelines. The LAMP assay was shown to be specific and extremely sensitive, because it detected FDp in all leaf samples that were determined to be FDp infected using quantitative real-time PCR. The whole procedure of sample preparation and testing was designed and optimized for on-site detection and can be completed in one hour. The homogenization procedure of the grapevine samples (leaf vein, flower or berry) was optimized to allow direct testing of crude homogenates with the LAMP assay, without the need for DNA extraction, and was shown to be extremely sensitive.
Keywords:flavescence dorée, homogenization, loop-mediated isothermal amplification, on-site application, validation
Publication status:Published
Publication version:Version of Record
Publication date:01.04.2015
Year of publishing:2015
Number of pages:str. 286-296
Numbering:Vol. 64, no. 2
PID:20.500.12556/DiRROS-19781 New window
UDC:577.2
ISSN on article:0032-0862
DOI:10.1111/ppa.12266 New window
COBISS.SI-ID:3175247 New window
Publication date in DiRROS:26.07.2024
Views:9
Downloads:4
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Record is a part of a journal

Title:Plant Pathology
Shortened title:Plant Pathol.
Publisher:Her Majesty's Stationery Office
ISSN:0032-0862
COBISS.SI-ID:5661191 New window

Document is financed by a project

Funder:EC - European Commission
Project number:262032
Name:COST-EFFECTIVE HAND-HELD DEVICE FOR RAPID IN-FIELD DETECTION OF FLAVENSCENCE DOREE PHYTOPLASMA IN GRAPEVINES
Acronym:VITISENS

Funder:EC - European Commission
Funding programme:Erasmus Student mobility for Placements 2011/2012
Name:scholarship

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License:CC BY-NC-ND 3.0, Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported
Link:http://creativecommons.org/licenses/by-nc-nd/3.0/
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