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Article

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

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Abiopep S.L. Parque Científico de Murcia. Ctra. Madrid Km 388, Complejo Espinardo. Edificio R 2ª Planta, Espinardo, 30100 Murcia, Spain
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Centro de Edafología y Biología Aplicada del Segura, Consejo Superior de Investigaciones Científicas (CSIC), Campus Universitario de Espinardo, Edificio 25, Espinardo, 30100 Murcia, Spain
*
Author to whom correspondence should be addressed.
Academic Editor: Gian Paolo Accotto
Viruses 2021, 13(9), 1680; https://doi.org/10.3390/v13091680
Received: 1 July 2021 / Revised: 13 August 2021 / Accepted: 23 August 2021 / Published: 25 August 2021
(This article belongs to the Section Viruses of Plants, Fungi and Protozoa)
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. View Full-Text
Keywords: ToBRFV; RT-qPCR; LAMP; DAS-ELISA; plant virus; tobamovirus; detection ToBRFV; RT-qPCR; LAMP; DAS-ELISA; plant virus; tobamovirus; detection
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MDPI and ACS Style

Bernabé-Orts, J.M.; Torre, C.; Méndez-López, E.; Hernando, Y.; Aranda, M.A. New Resources for the Specific and Sensitive Detection of the Emerging Tomato Brown Rugose Fruit Virus. Viruses 2021, 13, 1680. https://doi.org/10.3390/v13091680

AMA Style

Bernabé-Orts JM, Torre C, Méndez-López E, Hernando Y, Aranda MA. New Resources for the Specific and Sensitive Detection of the Emerging Tomato Brown Rugose Fruit Virus. Viruses. 2021; 13(9):1680. https://doi.org/10.3390/v13091680

Chicago/Turabian Style

Bernabé-Orts, Joan Miquel, Covadonga Torre, Eduardo Méndez-López, Yolanda Hernando, and Miguel A. Aranda. 2021. "New Resources for the Specific and Sensitive Detection of the Emerging Tomato Brown Rugose Fruit Virus" Viruses 13, no. 9: 1680. https://doi.org/10.3390/v13091680

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