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Real-time reverse transcription polymerase chain reaction development for rapid detection of Tomato brown rugose fruit virus and comparison with other techniques

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Real-time reverse transcription polymerase chain reaction development for rapid detection of Tomato brown rugose fruit virus and comparison with other techniques

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dc.contributor.author Panno, Stefano es_ES
dc.contributor.author Ruiz-Ruiz, Susana es_ES
dc.contributor.author Giovani Carusso, Andrea es_ES
dc.contributor.author Alfaro Fernández, Ana Olvido es_ES
dc.contributor.author Font San Ambrosio, Maria Isabel es_ES
dc.contributor.author Davino, Salvatore es_ES
dc.date.accessioned 2021-01-30T04:32:05Z
dc.date.available 2021-01-30T04:32:05Z
dc.date.issued 2019-10-17 es_ES
dc.identifier.uri http://hdl.handle.net/10251/160320
dc.description.abstract [EN] Background: Tomato brown rugose fruit virus (ToBRFV) is a highly infectious tobamovirus that causes severe disease in tomato (Solanum lycopersicum L.) crops. In Italy, the first ToBRFV outbreak occurred in 2018 in several provinces of the Sicily region. ToBRFV outbreak represents a serious threat for tomato crops in Italy and the Mediterranean Basin. Methods: Molecular and biological characterisation of the Sicilian ToBRFV ToB-SIC01/19 isolate was performed, and a sensitive and specific Real-time RT-PCR TaqMan minor groove binder probe method was developed to detect ToBRFV in infected plants and seeds. Moreover, four different sample preparation procedures (immunocapture, total RNA extraction, direct crude extract and leaf-disk crude extract) were evaluated. Results: The Sicilian isolate ToB-SIC01/19 (6,391 nt) showed a strong sequence identity with the isolates TBRFV-P12-3H and TBRFV-P12-3G from Germany, Tom 1-Jo from Jordan and TBRFV-IL from Israel. The ToB-SIC01/19 isolate was successfully transmitted by mechanical inoculations in S. lycopersicum L. and Capsicum annuum L., but no transmission occurred in S. melongena L. The developed real-time RT-PCR, based on the use of a primer set designed on conserved sequences in the open reading frames3, enabled a reliable quantitative detection. This method allowed clear discrimination of ToBRFV from other viruses belonging to the genus Tobamovirus, minimising false-negative results. Using immunocapture and total RNA extraction procedures, the real-time RT-PCR and end-point RT-PCR gave the same comparable results. Using direct crude extracts and leaf-disk crude extracts, the end-point RT-PCR was unable to provide a reliable result. This developed highly specific and sensitive real-time RT-PCR assay will be a particularly valuable tool for early ToBRFV diagnosis, optimising procedures in terms of costs and time. es_ES
dc.language Inglés es_ES
dc.publisher PeerJ es_ES
dc.relation.ispartof PeerJ es_ES
dc.rights Reconocimiento (by) es_ES
dc.subject ToBRFV es_ES
dc.subject RT-qPRC es_ES
dc.subject Fast detection es_ES
dc.subject.classification PRODUCCION VEGETAL es_ES
dc.title Real-time reverse transcription polymerase chain reaction development for rapid detection of Tomato brown rugose fruit virus and comparison with other techniques es_ES
dc.type Artículo es_ES
dc.identifier.doi 10.7717/peerj.7928 es_ES
dc.rights.accessRights Abierto es_ES
dc.contributor.affiliation Universitat Politècnica de València. Departamento de Ecosistemas Agroforestales - Departament d'Ecosistemes Agroforestals es_ES
dc.contributor.affiliation Universitat Politècnica de València. Instituto Agroforestal Mediterráneo - Institut Agroforestal Mediterrani es_ES
dc.description.bibliographicCitation Panno, S.; Ruiz-Ruiz, S.; Giovani Carusso, A.; Alfaro Fernández, AO.; Font San Ambrosio, MI.; Davino, S. (2019). Real-time reverse transcription polymerase chain reaction development for rapid detection of Tomato brown rugose fruit virus and comparison with other techniques. PeerJ. 7:1-20. https://doi.org/10.7717/peerj.7928 es_ES
dc.description.accrualMethod S es_ES
dc.relation.publisherversion https://doi.org/10.7717/peerj.7928 es_ES
dc.description.upvformatpinicio 1 es_ES
dc.description.upvformatpfin 20 es_ES
dc.type.version info:eu-repo/semantics/publishedVersion es_ES
dc.description.volume 7 es_ES
dc.identifier.eissn 2167-8359 es_ES
dc.identifier.pmid 31637144 es_ES
dc.identifier.pmcid PMC6800982 es_ES
dc.relation.pasarela S\407520 es_ES
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