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Detection, discrimination and absolute quantitation of Tomato spotted wilt virus isolates using real time RT-PCR with TaqMan®MGB probes

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Detection, discrimination and absolute quantitation of Tomato spotted wilt virus isolates using real time RT-PCR with TaqMan®MGB probes

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Nombre: -D.E. DEBRECZENI; ...
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dc.contributor.author Debreczeni, Diana Elvira es_ES
dc.contributor.author Ruiz Ruiz, Susana es_ES
dc.contributor.author Aramburu, J. es_ES
dc.contributor.author López Del Rincón, Carmelo es_ES
dc.contributor.author Belliure Ferrer, Belén Reyes es_ES
dc.contributor.author Galipienso Torregrosa, Luis es_ES
dc.contributor.author Soler Aleixandre, Salvador es_ES
dc.contributor.author Rubio, L. es_ES
dc.date.accessioned 2016-06-02T09:45:36Z
dc.date.available 2016-06-02T09:45:36Z
dc.date.issued 2011-09
dc.identifier.issn 0166-0934
dc.identifier.uri http://hdl.handle.net/10251/65093
dc.description.abstract A quantitative real-time reverse transcription-polymerase chain reaction (RT-qPCR) procedure using a general primer set and three TaqMan (R) MGB probes was developed for general and genotype-specific detection and quantitation of the genomic M segment of Tomato spotted wilt virus (TSWV). Standard curves using RNA transcripts homologous to the three probes allowed reproducible quantitative assays with a wide dynamic range (10(3)-10(10) TSWV M segment RNA copies/ng of total RNA) and high sensitivity. This protocol was assayed with a battery of TSWV isolates, covering the range of the present known genetic variation, in single and/or mix infections in three plant hosts, as well as in the thrips vector Frankliniella occidenialis. This quantitative detection assay will be a valuable tool for molecular biology and epidemiology studies, diagnosis and disease control. es_ES
dc.description.sponsorship This work was supported by grants INIA RTA2008-00010-C03 and Generalitat Valenciana ACOMP/2009/103 and ACOMP/2010/085. D.E. Debreczeni is the recipient of a FPU pre-doctoral fellowship from MEC and B. Belliure was supported by a INIA-CCAA contract. We thank I. Ferriol, Dr. S. Ambros and Dr. A. Olmos for technical advice; Dolores Comin for technical assistance; Dr. D. Peters and Dr. J. Contreras for kindly providing the TSWV isolate BR01 and a thrips colony, respectively; and Daniel R. Pearce for English revision. en_EN
dc.language Inglés es_ES
dc.publisher Elsevier es_ES
dc.relation.ispartof Journal of Virological Methods es_ES
dc.rights Reserva de todos los derechos es_ES
dc.subject TSWV es_ES
dc.subject Tospovirus es_ES
dc.subject Diagnosis es_ES
dc.subject Quantitative RT-PCR es_ES
dc.subject.classification GENETICA es_ES
dc.subject.classification MICROBIOLOGIA es_ES
dc.title Detection, discrimination and absolute quantitation of Tomato spotted wilt virus isolates using real time RT-PCR with TaqMan®MGB probes es_ES
dc.type Artículo es_ES
dc.identifier.doi 10.1016/j.jviromet.2011.05.027
dc.relation.projectID info:eu-repo/grantAgreement/MICINN//RTA2008-00010-C03-01/ES/Diversidad genética y factores evolutivos y epidemiológicos implicados en los aislados españoles de TSWV que superan las resistencias genéticas Sw-5 de tomate y Tsw de pimiento/ es_ES
dc.relation.projectID info:eu-repo/grantAgreement/Generalitat Valenciana//ACOMP%2F2009%2F103/ES/Diversidad genética y factores evolutivos y epidemiológicos implicados en los aislados españoles de TSWV que superan las resistencias genéticas Sw-5 de tomate y Tsw de pimiento/ es_ES
dc.rights.accessRights Cerrado es_ES
dc.contributor.affiliation Universitat Politècnica de València. Departamento de Biotecnología - Departament de Biotecnologia es_ES
dc.description.bibliographicCitation Debreczeni, DE.; Ruiz Ruiz, S.; Aramburu, J.; López Del Rincón, C.; Belliure Ferrer, BR.; Galipienso Torregrosa, L.; Soler Aleixandre, S.... (2011). Detection, discrimination and absolute quantitation of Tomato spotted wilt virus isolates using real time RT-PCR with TaqMan®MGB probes. Journal of Virological Methods. 176(1-2):32-37. https://doi.org/10.1016/j.jviromet.2011.05.027 es_ES
dc.description.accrualMethod S es_ES
dc.relation.publisherversion http://dx.doi.org/10.1016/j.jviromet.2011.05.027 es_ES
dc.description.upvformatpinicio 32 es_ES
dc.description.upvformatpfin 37 es_ES
dc.type.version info:eu-repo/semantics/publishedVersion es_ES
dc.description.volume 176 es_ES
dc.description.issue 1-2 es_ES
dc.relation.senia 200632 es_ES
dc.contributor.funder Generalitat Valenciana es_ES
dc.contributor.funder Ministerio de Ciencia e Innovación es_ES


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