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dc.contributor.authorBertolini, Edson
dc.contributor.authorOlmos, Antonio
dc.contributor.authorLópez, María M.
dc.contributor.authorCambra, Mariano
dc.date.accessioned2017-06-01T10:11:08Z
dc.date.available2017-06-01T10:11:08Z
dc.date.issued2003
dc.identifier.citationBertolini, E., Olmos, A., Lopez, M.M., Cambra, M. (2003). Multiplex nested reverse transcription-polymerase chain reaction in a single tube for sensitive and simultaneous detection of four RNA viruses and Pseudomonas savastanoi pv. savastanoi in olive trees. Phytopathology, 93(3), 286-292.
dc.identifier.issn0031-949X
dc.identifier.urihttp://hdl.handle.net/20.500.11939/4834
dc.description.abstractA multiplex nested reverse transcription-polymerase chain reaction (RT-PCR) in a single closed tube was developed for the simultaneous detection of four RNA viruses: Cucumber mosaic virus, Cherry leaf roll virus, Strawberry latent ringspot virus, and Arabis mosaic virus, and the bacterium Pseudomonas savastanoi pv. savastanoi. The method enabled, for the first time, the sensitive and simultaneous detection of RNA and DNA targets from plant viruses and a bacterium, saving time, decreasing risks of contamination, and reducing costs compared with conventional monospecific nested amplifications. The method was successfully coupled with colorimetric detection of amplicons using specific oligoprobes to simplify routine detection. Two hundred forty-five olive trees from 15 different cultivars were analyzed by multiplex RT-nested PCR coupled with colorimetric detection. Multiplex nested RT-PCR for viral detection increased the identification of positive trees by 8.1%. An uneven distribution of the viruses was observed in the infected trees. The bacterium was detected in 28.7% of the analyzed trees by the developed multiplex nested method and by a nested PCR previously developed. This powerful methodology could be applied to other models for the detection of several pathogens in a single assay.
dc.language.isoen
dc.titleMultiplex nested reverse transcription-polymerase chain reaction in a single tube for sensitive and simultaneous detection of four RNA viruses and Pseudomonas savastanoi pv. savastanoi in olive trees
dc.typearticle
dc.authorAddressInstituto Valenciano de Investigaciones Agrarias (IVIA), Carretera CV-315, Km. 10’7, 46113 Moncada (Valencia), Españaes
dc.date.issuedFreeFormMAR 2003
dc.entidadIVIACentro de Protección Vegetal y Biotecnología
dc.identifier.doi10.1094/PHYTO.2003.93.3.286
dc.journal.abbreviatedTitlePhytopathology
dc.journal.issueNumber3
dc.journal.titlePhytopathology
dc.journal.volumeNumber93
dc.page.final292
dc.page.initial286
dc.rights.accessRightsopenAccess
dc.source.typeImpreso


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