RT-PCR and dot blot hybridization methods for a universal detection of tospovirus.

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dc.contributorCNPH.
dc.creatorEIRAS, M.
dc.creatorRESENDE, R. O.
dc.creatorMISSIAGGIA. A. A.
dc.creatorAVILA, A. C. de
dc.date2024-10-21T20:53:56Z
dc.date2024-10-21T20:53:56Z
dc.date2001-07-20
dc.date2001
dc.date.accessioned2026-07-07T04:09:13Z
dc.descriptionTranscriptase reverse - polymerase chain reaction (RT-PCR) and dot blot hybridization with digoxigenin-labeled probes were applied for the universal detection of Tospovirus species. The virus species tested were Tomato spotted wilt virus, Tomato chlorotic spot virus, Groundnut ringspot virus, Chrysanthemum stem necrosis virus, Impatiens necrotic spot virus, Zucchini lethal chlorosis virus, Iris yellow spot virus. Primers for PCR amplification were designed to match conserved regions of the tospovirus genome. RT-PCR using distinct primer combinations was unable to simultaneously amplify all tospovirus species and consistently failed to detect ZLCV and IYSV in total RNA extracts. However, all tospovirus species were detected by RT-PCR when viral RNA was used as template. RNA-specific PCR products were used as probes for dot hybridization. This assay with a M probe (directed to the G1/G2 gene) detected at low stringency conditions all Tospovirus species, except IYSV. At low stringency conditions, the L non-radioactive probe detected the seven Tospovirus species in a single assay. This method for broad spectrum detection can be potentially employed in quarantine services for indexing in vitro germplasm.
dc.identifierFitopatologia Brasileira, Brasilia, v.26, n.2, p.170-175, jun. 2001.
dc.identifierhttp://www.alice.cnptia.embrapa.br/alice/handle/doc/769972
dc.identifierhttps://doi.org/10.1590/S0100-41582001000200009
dc.identifier.urihttp://hdl.handle.net/123456789/453187
dc.languageeng
dc.rightsopenAccess
dc.subjectHibridizacao
dc.subjectDot blot
dc.subjectDeteccao
dc.subjectRT-PCR
dc.subjectTospoviruses
dc.subjectTospovirus
dc.subjectdetection
dc.subjecthybridization
dc.titleRT-PCR and dot blot hybridization methods for a universal detection of tospovirus.
dc.typeArtigo de periódico

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