European protocol for diagnosis of Erwinia amylovora

The EU-funded project DIAGPRO has developed diagnostic protocols for 18 quarantine pests including Erwinia amylovora. The protocol for this organism includes a diagnosis scheme for detection of the bacterium in symptomatic and asymptomatic plants with details of the different techniques as isolation...

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Main Authors: López, María M., Llop, Pablo, Gorris, María T., Penyalver, Javier, Donat, Victoria, Cambra, Mariano, Keck, M.
Other Authors: Bazzi, C. Mazzucchi, U.
Format: Objeto de conferencia
Language:Inglés
Published: 2017
Online Access:http://hdl.handle.net/20.500.11939/5519
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author López, María M.
Llop, Pablo
Gorris, María T.
Penyalver, Javier
Donat, Victoria
Cambra, Mariano
Keck, M.
author2 Bazzi, C. Mazzucchi, U.
author_browse Bazzi, C. Mazzucchi, U.
Cambra, Mariano
Donat, Victoria
Gorris, María T.
Keck, M.
Llop, Pablo
López, María M.
Penyalver, Javier
author_facet Bazzi, C. Mazzucchi, U.
López, María M.
Llop, Pablo
Gorris, María T.
Penyalver, Javier
Donat, Victoria
Cambra, Mariano
Keck, M.
author_sort López, María M.
collection ReDivia
description The EU-funded project DIAGPRO has developed diagnostic protocols for 18 quarantine pests including Erwinia amylovora. The protocol for this organism includes a diagnosis scheme for detection of the bacterium in symptomatic and asymptomatic plants with details of the different techniques as isolation in three media, direct tissue print-ELISA, enrichment DASI-ELISA, immunofluorescence, conventional PCR and nested-PCR in a single closed tube (before and after enrichment of the samples in two liquid media). A ring test was conducted to evaluate and standardize some techniques and protocols. Testing was performed independently at 10 plant health laboratories analysing 15 samples (healthy, spiked or inoculated). The reliability of the diagnosis across all laboratories decreased with inoculum concentration. The accuracy of the different techniques, defined as (true positive diagnosis + true negatives)/total number of samples, was: 0.88, 0.92 and 0.92 respectively for isolation on King’s B, SNA and CCT media; 0.70, 0.72 and 0.66 for immunofluorescence performed with one monoclonal and two polyclonal antibodies commercially available; 0.79 and 0.83 for enrichment-ELISA with a commercial kit after enrichment in two media; 0.51 for conventional PCR and 0.69 for nested-PCR in a single tube, both techniques increasing their accuracy up to 0.78 and 0.86, respectively, after enrichment. The designed protocol is freely available at the web site: www.csl.gov.uk/science/organ/ph/diagpro.
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institution Instituto Valenciano de Investigaciones Agrarias (IVIA)
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spelling ReDivia55192025-04-25T14:53:26Z European protocol for diagnosis of Erwinia amylovora ACTA HORTICULTURAE López, María M. Llop, Pablo Gorris, María T. Penyalver, Javier Donat, Victoria Cambra, Mariano Keck, M. Bazzi, C. Mazzucchi, U. The EU-funded project DIAGPRO has developed diagnostic protocols for 18 quarantine pests including Erwinia amylovora. The protocol for this organism includes a diagnosis scheme for detection of the bacterium in symptomatic and asymptomatic plants with details of the different techniques as isolation in three media, direct tissue print-ELISA, enrichment DASI-ELISA, immunofluorescence, conventional PCR and nested-PCR in a single closed tube (before and after enrichment of the samples in two liquid media). A ring test was conducted to evaluate and standardize some techniques and protocols. Testing was performed independently at 10 plant health laboratories analysing 15 samples (healthy, spiked or inoculated). The reliability of the diagnosis across all laboratories decreased with inoculum concentration. The accuracy of the different techniques, defined as (true positive diagnosis + true negatives)/total number of samples, was: 0.88, 0.92 and 0.92 respectively for isolation on King’s B, SNA and CCT media; 0.70, 0.72 and 0.66 for immunofluorescence performed with one monoclonal and two polyclonal antibodies commercially available; 0.79 and 0.83 for enrichment-ELISA with a commercial kit after enrichment in two media; 0.51 for conventional PCR and 0.69 for nested-PCR in a single tube, both techniques increasing their accuracy up to 0.78 and 0.86, respectively, after enrichment. The designed protocol is freely available at the web site: www.csl.gov.uk/science/organ/ph/diagpro. 2017-06-01T10:12:30Z 2017-06-01T10:12:30Z 2006 2006 conferenceObject Lopez, M. M., Llop, P., Gorris, M. T., Penalver, J., Donat, V., Cambra, M. et al. (2006). European protocol for diagnosis of Erwinia amylovora. Proceedings of the Xth International Workshop on Fire Blight, (704), 99-103. 0567-7572; 90-6605-407-7 http://hdl.handle.net/20.500.11939/5519 10.17660/ActaHortic.2006.704.11 en openAccess Impreso
spellingShingle López, María M.
Llop, Pablo
Gorris, María T.
Penyalver, Javier
Donat, Victoria
Cambra, Mariano
Keck, M.
European protocol for diagnosis of Erwinia amylovora
title European protocol for diagnosis of Erwinia amylovora
title_full European protocol for diagnosis of Erwinia amylovora
title_fullStr European protocol for diagnosis of Erwinia amylovora
title_full_unstemmed European protocol for diagnosis of Erwinia amylovora
title_short European protocol for diagnosis of Erwinia amylovora
title_sort european protocol for diagnosis of erwinia amylovora
url http://hdl.handle.net/20.500.11939/5519
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