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Article

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

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CREA—Research Centre for Plant Protection and Certification, Via C.G. Bertero 22, 00156 Roma, Italy
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Department of Biotechnology and Systems Biology, National Institute of Biology, Večna pot 111, SI-1000 Ljubljana, Slovenia
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School for Viticulture and Enology, University of Nova Gorica, Dvorec Lanthieri, Glavni trg 8, SI-5271 Vipava, Slovenia
*
Author to whom correspondence should be addressed.
Academic Editors: Aviv Dombrovsky and Ozgur Batuman
Plants 2022, 11(4), 489; https://doi.org/10.3390/plants11040489
Received: 30 December 2021 / Revised: 31 January 2022 / Accepted: 9 February 2022 / Published: 11 February 2022
(This article belongs to the Special Issue Tobamoviruses and Interacting Viruses in Modern Agriculture)
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. View Full-Text
Keywords: ToBRFV; ToMMV; Solanum lycopersicum; Capsicum annum; leaves detection; seeds detections; performance criteria ToBRFV; ToMMV; Solanum lycopersicum; Capsicum annum; leaves detection; seeds detections; performance criteria
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MDPI and ACS Style

Tiberini, A.; Manglli, A.; Taglienti, A.; Vučurović, A.; Brodarič, J.; Ferretti, L.; Luigi, M.; Gentili, A.; Mehle, N. 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. Plants 2022, 11, 489. https://doi.org/10.3390/plants11040489

AMA Style

Tiberini A, Manglli A, Taglienti A, Vučurović A, Brodarič J, Ferretti L, Luigi M, Gentili A, Mehle N. 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. Plants. 2022; 11(4):489. https://doi.org/10.3390/plants11040489

Chicago/Turabian Style

Tiberini, Antonio, Ariana Manglli, Anna Taglienti, Ana Vučurović, Jakob Brodarič, Luca Ferretti, Marta Luigi, Andrea Gentili, and Nataša Mehle. 2022. "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" Plants 11, no. 4: 489. https://doi.org/10.3390/plants11040489

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