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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 principales: Bernabé-Orts, Joan Miquel, Torre, Covadonga, Méndez-López, Eduardo, Hernando, Yolanda, Aranda, Miguel A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8473139/
https://www.ncbi.nlm.nih.gov/pubmed/34578261
http://dx.doi.org/10.3390/v13091680
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author Bernabé-Orts, Joan Miquel
Torre, Covadonga
Méndez-López, Eduardo
Hernando, Yolanda
Aranda, Miguel A.
author_facet Bernabé-Orts, Joan Miquel
Torre, Covadonga
Méndez-López, Eduardo
Hernando, Yolanda
Aranda, Miguel A.
author_sort Bernabé-Orts, Joan Miquel
collection PubMed
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.
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spelling pubmed-84731392021-09-28 New Resources for the Specific and Sensitive Detection of the Emerging Tomato Brown Rugose Fruit Virus Bernabé-Orts, Joan Miquel Torre, Covadonga Méndez-López, Eduardo Hernando, Yolanda Aranda, Miguel A. Viruses Article 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. MDPI 2021-08-25 /pmc/articles/PMC8473139/ /pubmed/34578261 http://dx.doi.org/10.3390/v13091680 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Bernabé-Orts, Joan Miquel
Torre, Covadonga
Méndez-López, Eduardo
Hernando, Yolanda
Aranda, Miguel A.
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
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_short 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
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8473139/
https://www.ncbi.nlm.nih.gov/pubmed/34578261
http://dx.doi.org/10.3390/v13091680
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