Real-time RT-PCR for quantitative detection of Plum pox virus
Two protocols for real-time quantitative detection of Plum pox virus (PPV) and identification of PPV-D and PPV-M isolates were developed using the intercalating dye SYBR-Green I and TaqMan chemistry. SYBR-Green I method using the universal PPV primers, P1 and P2, required for its optimization an acc...
| Autores principales: | , , , |
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| Otros Autores: | |
| Formato: | conferenceObject |
| Lenguaje: | Inglés |
| Publicado: |
2017
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| Acceso en línea: | http://hdl.handle.net/20.500.11939/4257 |
| _version_ | 1855032090315194368 |
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| author | Olmos, Antonio Bertolini, Edson Gil, Maite Cambra, Mariano |
| author2 | Llácer, Gerardo |
| author_browse | Bertolini, Edson Cambra, Mariano Gil, Maite Llácer, Gerardo Olmos, Antonio |
| author_facet | Llácer, Gerardo Olmos, Antonio Bertolini, Edson Gil, Maite Cambra, Mariano |
| author_sort | Olmos, Antonio |
| collection | ReDivia |
| description | Two protocols for real-time quantitative detection of Plum pox virus (PPV) and identification of PPV-D and PPV-M isolates were developed using the intercalating dye SYBR-Green I and TaqMan chemistry. SYBR-Green I method using the universal PPV primers, P1 and P2, required for its optimization an accurate analysis of primers concentration to avoid primer-dimer formation. Optimization for TaqMan technology required the design of new primers and internal probes using the Primer Express software and the alignment of available PPV sequences from Genbank, EMBL, and DDBJ databases. Although the two methods were able to detect as low as 4 copies of transcripts, only the TaqMan method maintained this sensitivity when plant material was used. In addition, the use of TaqMan technology succeed in the detection and quantification of PPV targets in single aphids. |
| format | conferenceObject |
| id | ReDivia4257 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia42572025-04-25T14:51:40Z Real-time RT-PCR for quantitative detection of Plum pox virus ACTA HORTICULTURAE Olmos, Antonio Bertolini, Edson Gil, Maite Cambra, Mariano Llácer, Gerardo Two protocols for real-time quantitative detection of Plum pox virus (PPV) and identification of PPV-D and PPV-M isolates were developed using the intercalating dye SYBR-Green I and TaqMan chemistry. SYBR-Green I method using the universal PPV primers, P1 and P2, required for its optimization an accurate analysis of primers concentration to avoid primer-dimer formation. Optimization for TaqMan technology required the design of new primers and internal probes using the Primer Express software and the alignment of available PPV sequences from Genbank, EMBL, and DDBJ databases. Although the two methods were able to detect as low as 4 copies of transcripts, only the TaqMan method maintained this sensitivity when plant material was used. In addition, the use of TaqMan technology succeed in the detection and quantification of PPV targets in single aphids. 2017-06-01T10:09:42Z 2017-06-01T10:09:42Z 2004 2004 conferenceObject Olmos, A., Bertolini, E., Gil, M., Cambra, M. (2004). Real-time RT-PCR for quantitative detection of Plum pox virus. Proceedings of the Xixth International Symposium on Virus and Virus-Like Diseases of Temperate Fruit Crops: Fruit Tree Diseases, (657), 149-153. 0567-7572; 90-6605-148-5 http://hdl.handle.net/20.500.11939/4257 10.17660/ActaHortic.2004.657.19 en openAccess Impreso |
| spellingShingle | Olmos, Antonio Bertolini, Edson Gil, Maite Cambra, Mariano Real-time RT-PCR for quantitative detection of Plum pox virus |
| title | Real-time RT-PCR for quantitative detection of Plum pox virus |
| title_full | Real-time RT-PCR for quantitative detection of Plum pox virus |
| title_fullStr | Real-time RT-PCR for quantitative detection of Plum pox virus |
| title_full_unstemmed | Real-time RT-PCR for quantitative detection of Plum pox virus |
| title_short | Real-time RT-PCR for quantitative detection of Plum pox virus |
| title_sort | real time rt pcr for quantitative detection of plum pox virus |
| url | http://hdl.handle.net/20.500.11939/4257 |
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