New Resources for the Specific and Sensitive Detection of the Emerging Tomato Brown Rugose Fruit Virus

Plant viruses can evolve towards new pathogenic entities that may eventually cause outbreaks and become epidemics or even pandemics. Seven years ago, tomato brown rugose fruit virus (ToBRFV) emerged, overcoming the genetic resistance that had been employed for more than sixty years against tobamovir...

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Autores: Bernabé-Orts, Joan Miquel, Torre, Covadonga, Méndez-López, Eduardo, Hernando Saiz, Yolanda, Aranda, Miguel A.
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2021
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/251439
Acceso en línea:http://hdl.handle.net/10261/251439
Access Level:acceso abierto
Palabra clave:ToBRFV
RT-qPCR
LAMP
DAS-ELISA
Plant virus
Tobamovirus
Detection
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spelling New Resources for the Specific and Sensitive Detection of the Emerging Tomato Brown Rugose Fruit VirusBernabé-Orts, Joan MiquelTorre, CovadongaMéndez-López, EduardoHernando Saiz, YolandaAranda, Miguel A.ToBRFVRT-qPCRLAMPDAS-ELISAPlant virusTobamovirusDetectionPlant viruses can evolve towards new pathogenic entities that may eventually cause outbreaks and become epidemics or even pandemics. Seven years ago, tomato brown rugose fruit virus (ToBRFV) emerged, overcoming the genetic resistance that had been employed for more than sixty years against tobamoviruses in tomato. Since then, ToBRFV has spread worldwide, producing significant losses in tomato crops. While new resistances are deployed, the only means of control is the implementation of effective prevention and eradication strategies. For this purpose, in this work, we have designed, assessed, and compared an array of tests for the specific and sensitive detection of the ToBRFV in leaf samples. First, two monoclonal antibodies were generated against a singular peptide of the ToBRFV coat protein; antibodies were utilized to devise a double-antibody-sandwich enzyme-linked immunosorbent assay (DAS-ELISA) test that sensitively detects this virus and has no cross-reactivity with other related tobamoviruses. Second, a real-time quantitative PCR (RT-qPCR) test targeting the RNA-dependent replicase open reading frame (ORF) was designed, and its performance and specificity validated in comparison with the CaTa28 and CSP1325 tests recommended by plant protection authorities in Europe. Third, in line with the tendency to use field-deployable diagnostic techniques, we developed and tested two sets of loop-mediated isothermal amplification (LAMP) primers to double-check the detection of the movement protein ORF of ToBRFV, and one set that works as an internal control. Finally, we compared all of these methods by employing a collection of samples with different ToBRFV loads to evaluate the overall performance of each test.Work at the CEBAS-CSIC lab was partially supported by grant RTI2018-097099-B-I00 from the Spanish Ministry of Science and Innovation. Work at Abiopep was partially supported by grant PTQ-2019-010500.Peer reviewedMultidisciplinary Digital Publishing InstituteMinisterio de Ciencia, Innovación y Universidades (España)Agencia Estatal de Investigación (España)Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2021202120212021info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/251439reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-097099-B-I00info:eu-repo/grantAgreement/AEI//PTQ-2019-010500Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2514392026-05-22T06:33:51Z
dc.title.none.fl_str_mv New Resources for the Specific and Sensitive Detection of the Emerging Tomato Brown Rugose Fruit Virus
title New Resources for the Specific and Sensitive Detection of the Emerging Tomato Brown Rugose Fruit Virus
spellingShingle New Resources for the Specific and Sensitive Detection of the Emerging Tomato Brown Rugose Fruit Virus
Bernabé-Orts, Joan Miquel
ToBRFV
RT-qPCR
LAMP
DAS-ELISA
Plant virus
Tobamovirus
Detection
title_short New Resources for the Specific and Sensitive Detection of the Emerging Tomato Brown Rugose Fruit Virus
title_full New Resources for the Specific and Sensitive Detection of the Emerging Tomato Brown Rugose Fruit Virus
title_fullStr New Resources for the Specific and Sensitive Detection of the Emerging Tomato Brown Rugose Fruit Virus
title_full_unstemmed New Resources for the Specific and Sensitive Detection of the Emerging Tomato Brown Rugose Fruit Virus
title_sort New Resources for the Specific and Sensitive Detection of the Emerging Tomato Brown Rugose Fruit Virus
dc.creator.none.fl_str_mv Bernabé-Orts, Joan Miquel
Torre, Covadonga
Méndez-López, Eduardo
Hernando Saiz, Yolanda
Aranda, Miguel A.
author Bernabé-Orts, Joan Miquel
author_facet Bernabé-Orts, Joan Miquel
Torre, Covadonga
Méndez-López, Eduardo
Hernando Saiz, Yolanda
Aranda, Miguel A.
author_role author
author2 Torre, Covadonga
Méndez-López, Eduardo
Hernando Saiz, Yolanda
Aranda, Miguel A.
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Ciencia, Innovación y Universidades (España)
Agencia Estatal de Investigación (España)
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv ToBRFV
RT-qPCR
LAMP
DAS-ELISA
Plant virus
Tobamovirus
Detection
topic ToBRFV
RT-qPCR
LAMP
DAS-ELISA
Plant virus
Tobamovirus
Detection
description Plant viruses can evolve towards new pathogenic entities that may eventually cause outbreaks and become epidemics or even pandemics. Seven years ago, tomato brown rugose fruit virus (ToBRFV) emerged, overcoming the genetic resistance that had been employed for more than sixty years against tobamoviruses in tomato. Since then, ToBRFV has spread worldwide, producing significant losses in tomato crops. While new resistances are deployed, the only means of control is the implementation of effective prevention and eradication strategies. For this purpose, in this work, we have designed, assessed, and compared an array of tests for the specific and sensitive detection of the ToBRFV in leaf samples. First, two monoclonal antibodies were generated against a singular peptide of the ToBRFV coat protein; antibodies were utilized to devise a double-antibody-sandwich enzyme-linked immunosorbent assay (DAS-ELISA) test that sensitively detects this virus and has no cross-reactivity with other related tobamoviruses. Second, a real-time quantitative PCR (RT-qPCR) test targeting the RNA-dependent replicase open reading frame (ORF) was designed, and its performance and specificity validated in comparison with the CaTa28 and CSP1325 tests recommended by plant protection authorities in Europe. Third, in line with the tendency to use field-deployable diagnostic techniques, we developed and tested two sets of loop-mediated isothermal amplification (LAMP) primers to double-check the detection of the movement protein ORF of ToBRFV, and one set that works as an internal control. Finally, we compared all of these methods by employing a collection of samples with different ToBRFV loads to evaluate the overall performance of each test.
publishDate 2021
dc.date.none.fl_str_mv 2021
2021
2021
2021
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/251439
url http://hdl.handle.net/10261/251439
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-097099-B-I00
info:eu-repo/grantAgreement/AEI//PTQ-2019-010500
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
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repository.mail.fl_str_mv
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