Detection, discrimination and absolute quantitation of Tomato spotted wilt virus isolates using real time RT-PCR with TaqMan (R) MGB probes
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 cu...
| Main Authors: | , , , , , , , |
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| Format: | article |
| Language: | Inglés |
| Published: |
2017
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| Online Access: | http://hdl.handle.net/20.500.11939/5111 |
| _version_ | 1855032227366174720 |
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| author | Debreczeni, Diana E. Ruiz-Ruiz, Susana Aramburu, Jose Lopez, Carmelo Belliure, Belén Galipienso, Luis Soler, Salvador Rubio, Luis |
| author_browse | Aramburu, Jose Belliure, Belén Debreczeni, Diana E. Galipienso, Luis Lopez, Carmelo Rubio, Luis Ruiz-Ruiz, Susana Soler, Salvador |
| author_facet | Debreczeni, Diana E. Ruiz-Ruiz, Susana Aramburu, Jose Lopez, Carmelo Belliure, Belén Galipienso, Luis Soler, Salvador Rubio, Luis |
| author_sort | Debreczeni, Diana E. |
| collection | ReDivia |
| description | 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. |
| format | article |
| id | ReDivia5111 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia51112025-04-25T14:45:24Z Detection, discrimination and absolute quantitation of Tomato spotted wilt virus isolates using real time RT-PCR with TaqMan (R) MGB probes Debreczeni, Diana E. Ruiz-Ruiz, Susana Aramburu, Jose Lopez, Carmelo Belliure, Belén Galipienso, Luis Soler, Salvador Rubio, Luis 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. 2017-06-01T10:11:43Z 2017-06-01T10:11:43Z 2011 SEP 2011 article Debreczeni, D. E., Ruiz-Ruiz, S., Aramburu, J., Lopez, C., Belliure, B., Galipienso, L., Soler, S., Rubio, L. (2011). Detection, discrimination and absolute quantitation of Tomato spotted wilt virus isolates using real time RT-PCR with TaqMan (R), MGB probes. Journal of virological methods, 176(1-2), 32-37. 0166-0934 http://hdl.handle.net/20.500.11939/5111 10.1016/j.jviromet.2011.05.027 en openAccess Impreso |
| spellingShingle | Debreczeni, Diana E. Ruiz-Ruiz, Susana Aramburu, Jose Lopez, Carmelo Belliure, Belén Galipienso, Luis Soler, Salvador Rubio, Luis Detection, discrimination and absolute quantitation of Tomato spotted wilt virus isolates using real time RT-PCR with TaqMan (R) MGB probes |
| title | Detection, discrimination and absolute quantitation of Tomato spotted wilt virus isolates using real time RT-PCR with TaqMan (R) MGB probes |
| title_full | Detection, discrimination and absolute quantitation of Tomato spotted wilt virus isolates using real time RT-PCR with TaqMan (R) MGB probes |
| title_fullStr | Detection, discrimination and absolute quantitation of Tomato spotted wilt virus isolates using real time RT-PCR with TaqMan (R) MGB probes |
| title_full_unstemmed | Detection, discrimination and absolute quantitation of Tomato spotted wilt virus isolates using real time RT-PCR with TaqMan (R) MGB probes |
| title_short | Detection, discrimination and absolute quantitation of Tomato spotted wilt virus isolates using real time RT-PCR with TaqMan (R) MGB probes |
| title_sort | detection discrimination and absolute quantitation of tomato spotted wilt virus isolates using real time rt pcr with taqman r mgb probes |
| url | http://hdl.handle.net/20.500.11939/5111 |
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