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
Por favor, use este identificador para citar o enlazar este ítem: http://hdl.handle.net/10251/65093
Title:
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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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Author:
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Debreczeni, Diana Elvira
Ruiz Ruiz, Susana
Aramburu, J.
López Del Rincón, Carmelo
Belliure Ferrer, Belén Reyes
Galipienso Torregrosa, Luis
Soler Aleixandre, Salvador
Rubio, L.
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UPV Unit:
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Universitat Politècnica de València. Departamento de Biotecnología - Departament de Biotecnologia
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Issued date:
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Abstract:
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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 ...[+]
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.
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Subjects:
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TSWV
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Tospovirus
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Diagnosis
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Quantitative RT-PCR
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Copyrigths:
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Cerrado |
Source:
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Journal of Virological Methods. (issn:
0166-0934
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DOI:
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10.1016/j.jviromet.2011.05.027
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Publisher:
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Elsevier
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Publisher version:
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http://dx.doi.org/10.1016/j.jviromet.2011.05.027
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Project ID:
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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/
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/
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Thanks:
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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 ...[+]
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.
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Type:
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Artículo
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