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Development and Validation of a One-Step Reverse Transcription Real-Time PCR Assay for Simultaneous Detection and Identification of Tomato Mottle Mosaic Virus and Tomato Brown Rugose Fruit Virus

Tobamovirus species represent a threat to solanaceous crops worldwide, due to their extreme stability and because they are seed borne. In particular, recent outbreaks of tomato brown rugose fruit virus in tomato and pepper crops led to the establishment of prompt control measures, and the need for r...

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Autores principales: Tiberini, Antonio, Manglli, Ariana, Taglienti, Anna, Vučurović, Ana, Brodarič, Jakob, Ferretti, Luca, Luigi, Marta, Gentili, Andrea, Mehle, Nataša
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8878898/
https://www.ncbi.nlm.nih.gov/pubmed/35214821
http://dx.doi.org/10.3390/plants11040489
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author Tiberini, Antonio
Manglli, Ariana
Taglienti, Anna
Vučurović, Ana
Brodarič, Jakob
Ferretti, Luca
Luigi, Marta
Gentili, Andrea
Mehle, Nataša
author_facet Tiberini, Antonio
Manglli, Ariana
Taglienti, Anna
Vučurović, Ana
Brodarič, Jakob
Ferretti, Luca
Luigi, Marta
Gentili, Andrea
Mehle, Nataša
author_sort Tiberini, Antonio
collection PubMed
description Tobamovirus species represent a threat to solanaceous crops worldwide, due to their extreme stability and because they are seed borne. In particular, recent outbreaks of tomato brown rugose fruit virus in tomato and pepper crops led to the establishment of prompt control measures, and the need for reliable diagnosis was urged. Another member of the genus, tomato mottle mosaic virus, has recently gained attention due to reports in different continents and its common features with tomato brown rugose fruit virus. In this study, a new real-time RT-PCR detection system was developed for tomato brown rugose fruit virus and tomato mottle mosaic virus on tomato leaves and seeds using TaqMan chemistry. This test was designed to detect tomato mottle mosaic virus by amplifying the movement protein gene in a duplex assay with the tomato brown rugose fruit virus target on the CP-3’NTR region, which was previously validated as a single assay. The performance of this test was evaluated, displaying analytical sensitivity 10(−5)–10(−6)-fold dilution for seeds and leaves, respectively, and good analytical specificity, repeatability, and reproducibility. Using the newly developed and validated test, tomato brown rugose fruit virus detection was 100% concordant with previously performed analyses on 106 official samples collected in 2021 from different continents.
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spelling pubmed-88788982022-02-26 Development and Validation of a One-Step Reverse Transcription Real-Time PCR Assay for Simultaneous Detection and Identification of Tomato Mottle Mosaic Virus and Tomato Brown Rugose Fruit Virus Tiberini, Antonio Manglli, Ariana Taglienti, Anna Vučurović, Ana Brodarič, Jakob Ferretti, Luca Luigi, Marta Gentili, Andrea Mehle, Nataša Plants (Basel) Article Tobamovirus species represent a threat to solanaceous crops worldwide, due to their extreme stability and because they are seed borne. In particular, recent outbreaks of tomato brown rugose fruit virus in tomato and pepper crops led to the establishment of prompt control measures, and the need for reliable diagnosis was urged. Another member of the genus, tomato mottle mosaic virus, has recently gained attention due to reports in different continents and its common features with tomato brown rugose fruit virus. In this study, a new real-time RT-PCR detection system was developed for tomato brown rugose fruit virus and tomato mottle mosaic virus on tomato leaves and seeds using TaqMan chemistry. This test was designed to detect tomato mottle mosaic virus by amplifying the movement protein gene in a duplex assay with the tomato brown rugose fruit virus target on the CP-3’NTR region, which was previously validated as a single assay. The performance of this test was evaluated, displaying analytical sensitivity 10(−5)–10(−6)-fold dilution for seeds and leaves, respectively, and good analytical specificity, repeatability, and reproducibility. Using the newly developed and validated test, tomato brown rugose fruit virus detection was 100% concordant with previously performed analyses on 106 official samples collected in 2021 from different continents. MDPI 2022-02-11 /pmc/articles/PMC8878898/ /pubmed/35214821 http://dx.doi.org/10.3390/plants11040489 Text en © 2022 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
Tiberini, Antonio
Manglli, Ariana
Taglienti, Anna
Vučurović, Ana
Brodarič, Jakob
Ferretti, Luca
Luigi, Marta
Gentili, Andrea
Mehle, Nataša
Development and Validation of a One-Step Reverse Transcription Real-Time PCR Assay for Simultaneous Detection and Identification of Tomato Mottle Mosaic Virus and Tomato Brown Rugose Fruit Virus
title Development and Validation of a One-Step Reverse Transcription Real-Time PCR Assay for Simultaneous Detection and Identification of Tomato Mottle Mosaic Virus and Tomato Brown Rugose Fruit Virus
title_full Development and Validation of a One-Step Reverse Transcription Real-Time PCR Assay for Simultaneous Detection and Identification of Tomato Mottle Mosaic Virus and Tomato Brown Rugose Fruit Virus
title_fullStr Development and Validation of a One-Step Reverse Transcription Real-Time PCR Assay for Simultaneous Detection and Identification of Tomato Mottle Mosaic Virus and Tomato Brown Rugose Fruit Virus
title_full_unstemmed Development and Validation of a One-Step Reverse Transcription Real-Time PCR Assay for Simultaneous Detection and Identification of Tomato Mottle Mosaic Virus and Tomato Brown Rugose Fruit Virus
title_short Development and Validation of a One-Step Reverse Transcription Real-Time PCR Assay for Simultaneous Detection and Identification of Tomato Mottle Mosaic Virus and Tomato Brown Rugose Fruit Virus
title_sort development and validation of a one-step reverse transcription real-time pcr assay for simultaneous detection and identification of tomato mottle mosaic virus and tomato brown rugose fruit virus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8878898/
https://www.ncbi.nlm.nih.gov/pubmed/35214821
http://dx.doi.org/10.3390/plants11040489
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