European Diagnostic Protocols (DIAGPRO) for Citrus tristeza virus in Adult Trees

DIAGPRO is a European Union (EU)-funded project to develop and validate European schemes for detection and diagnosis of a broad range of important organisms harmful to plants, according the EU Plant Health Directive (2000/29 as amended). Citrus tristeza virus (CTV) is included, and here are reported...

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Autores principales: Cambra, Mariano, Gorris, María T., Olmos, Antonio, Martínez, M. Carmen, Roman, M. P., Bertolini, Edson, Lopez, A., Carbonell, Emilio A.
Otros Autores: Durán-Vila, Núria
Formato: conferenceObject
Lenguaje:Inglés
Publicado: IOCV 2021
Materias:
Acceso en línea:http://hdl.handle.net/20.500.11939/7791
https://iocv.ucr.edu/sites/g/files/rcwecm4696/files/2020-06/069-77%20IOCV15.pdf
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author Cambra, Mariano
Gorris, María T.
Olmos, Antonio
Martínez, M. Carmen
Roman, M. P.
Bertolini, Edson
Lopez, A.
Carbonell, Emilio A.
author2 Durán-Vila, Núria
author_browse Bertolini, Edson
Cambra, Mariano
Carbonell, Emilio A.
Durán-Vila, Núria
Gorris, María T.
Lopez, A.
Martínez, M. Carmen
Olmos, Antonio
Roman, M. P.
author_facet Durán-Vila, Núria
Cambra, Mariano
Gorris, María T.
Olmos, Antonio
Martínez, M. Carmen
Roman, M. P.
Bertolini, Edson
Lopez, A.
Carbonell, Emilio A.
author_sort Cambra, Mariano
collection ReDivia
description DIAGPRO is a European Union (EU)-funded project to develop and validate European schemes for detection and diagnosis of a broad range of important organisms harmful to plants, according the EU Plant Health Directive (2000/29 as amended). Citrus tristeza virus (CTV) is included, and here are reported the results of a study to compare and standardize diagnostic methods for this virus. The most appropriate protocols were found to be graft inoculation of Mexican lime, sweet orange and grapefruit seedlings, DAS and DASI-ELISA, tissue print-ELISA, immunocapture RT-PCR and immunocapture nested RT-PCR in a single closed tube (conventional, print and squash capture formats). Samples of adult trees were tested during the different seasons (springtime gives the highest CTV titers) using five young shoots or fruit peduncles or 10 fullyexpanded leaves. Five flowers or fruits per tree, when available, were also found appropriate for reliable detection in recently infected adult trees. Different commercial polyclonal antibodies and the mixture of monoclonal antibodies (MCAs) 3DF1 and 3CA5 are recommended after calibration for conventional ELISA; the same MCAs are recommended for tissue print ELISA, and the primers of Olmos et al., 1999 (NAR 27, 1564-1565) for RT-PCR variants. A combination of two selected methods based on biological indexing, serological or molecular detection, will be required to officially declare a positive detection. Tissue print-ELISA using specific MCAs was found to be the most reliable, simple and economic method for routine analysis of plant material. The protocols and reagents have been ring tested and are published at http://www.csl.gov.uk/prodserve/know/ diagpro/ctv.pdf, an Internet website for comments before publication in the EU Official Bulletin.
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spelling ReDivia77912025-04-25T14:54:08Z European Diagnostic Protocols (DIAGPRO) for Citrus tristeza virus in Adult Trees Cambra, Mariano Gorris, María T. Olmos, Antonio Martínez, M. Carmen Roman, M. P. Bertolini, Edson Lopez, A. Carbonell, Emilio A. Durán-Vila, Núria Detection H20 Plant diseases H01 Protection of plants - General aspects U30 Research methods protocols Diagnosis ELISA Plant health PCR Citrus tristeza virus DIAGPRO is a European Union (EU)-funded project to develop and validate European schemes for detection and diagnosis of a broad range of important organisms harmful to plants, according the EU Plant Health Directive (2000/29 as amended). Citrus tristeza virus (CTV) is included, and here are reported the results of a study to compare and standardize diagnostic methods for this virus. The most appropriate protocols were found to be graft inoculation of Mexican lime, sweet orange and grapefruit seedlings, DAS and DASI-ELISA, tissue print-ELISA, immunocapture RT-PCR and immunocapture nested RT-PCR in a single closed tube (conventional, print and squash capture formats). Samples of adult trees were tested during the different seasons (springtime gives the highest CTV titers) using five young shoots or fruit peduncles or 10 fullyexpanded leaves. Five flowers or fruits per tree, when available, were also found appropriate for reliable detection in recently infected adult trees. Different commercial polyclonal antibodies and the mixture of monoclonal antibodies (MCAs) 3DF1 and 3CA5 are recommended after calibration for conventional ELISA; the same MCAs are recommended for tissue print ELISA, and the primers of Olmos et al., 1999 (NAR 27, 1564-1565) for RT-PCR variants. A combination of two selected methods based on biological indexing, serological or molecular detection, will be required to officially declare a positive detection. Tissue print-ELISA using specific MCAs was found to be the most reliable, simple and economic method for routine analysis of plant material. The protocols and reagents have been ring tested and are published at http://www.csl.gov.uk/prodserve/know/ diagpro/ctv.pdf, an Internet website for comments before publication in the EU Official Bulletin. 2021-11-30T09:16:14Z 2021-11-30T09:16:14Z 2002 conferenceObject Cambra, M., Gorris, M. T., Olmos, A., Martínez, M. C., Román, M. P., Bertolini, E. et al. (2002). European diagnostic protocols (DIAGPRO) for Citrus tristeza virus in adult trees. Proceedings of the Fifteenth Conference of the International Organization of Citrus Virologists, 69-75. 2313-5123 http://hdl.handle.net/20.500.11939/7791 https://iocv.ucr.edu/sites/g/files/rcwecm4696/files/2020-06/069-77%20IOCV15.pdf en 2001-11-11 Fifteenth Conference of the International Organization of Citrus Virologists Paphos, Cyprus Financial support was provided by the European Commission, through the SMT program project SMT4-CT98-2252 (DIAGPRO) and by grants from INIA (SC 98-060) and Consellería de Agricultura, Pesca y Alimentación de la Generalidad Valenciana (IVIA grant 5901). Atribución-NoComercial-SinDerivadas 3.0 España http://creativecommons.org/licenses/by-nc-nd/3.0/es/ openAccess IOCV electronico
spellingShingle Detection
H20 Plant diseases
H01 Protection of plants - General aspects
U30 Research methods
protocols
Diagnosis
ELISA
Plant health
PCR
Citrus tristeza virus
Cambra, Mariano
Gorris, María T.
Olmos, Antonio
Martínez, M. Carmen
Roman, M. P.
Bertolini, Edson
Lopez, A.
Carbonell, Emilio A.
European Diagnostic Protocols (DIAGPRO) for Citrus tristeza virus in Adult Trees
title European Diagnostic Protocols (DIAGPRO) for Citrus tristeza virus in Adult Trees
title_full European Diagnostic Protocols (DIAGPRO) for Citrus tristeza virus in Adult Trees
title_fullStr European Diagnostic Protocols (DIAGPRO) for Citrus tristeza virus in Adult Trees
title_full_unstemmed European Diagnostic Protocols (DIAGPRO) for Citrus tristeza virus in Adult Trees
title_short European Diagnostic Protocols (DIAGPRO) for Citrus tristeza virus in Adult Trees
title_sort european diagnostic protocols diagpro for citrus tristeza virus in adult trees
topic Detection
H20 Plant diseases
H01 Protection of plants - General aspects
U30 Research methods
protocols
Diagnosis
ELISA
Plant health
PCR
Citrus tristeza virus
url http://hdl.handle.net/20.500.11939/7791
https://iocv.ucr.edu/sites/g/files/rcwecm4696/files/2020-06/069-77%20IOCV15.pdf
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