Detection and quantitation of Citrus leaf blotch virus by TaqMan real-time RT-PCR
A real-time RT-PCR assay based on the TaqMan chemistry was developed for reliable detection and quantitation of Citrus leaf blotch virus (CLBV) in citrus plants. Detection by this method was highly specific and about one thousand times more sensitive than detection by conventional RT-PCR. An externa...
| Autores principales: | , , , , , |
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| Formato: | Artículo |
| Lenguaje: | Inglés |
| Publicado: |
2017
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| Acceso en línea: | http://hdl.handle.net/20.500.11939/4456 |
| _version_ | 1855491757175734272 |
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| author | Ruiz-Ruiz, Susana Ambrós, Silvia Vives, María C. Navarro, Luis Moreno, Pedro Guerri, José |
| author_browse | Ambrós, Silvia Guerri, José Moreno, Pedro Navarro, Luis Ruiz-Ruiz, Susana Vives, María C. |
| author_facet | Ruiz-Ruiz, Susana Ambrós, Silvia Vives, María C. Navarro, Luis Moreno, Pedro Guerri, José |
| author_sort | Ruiz-Ruiz, Susana |
| collection | ReDivia |
| description | A real-time RT-PCR assay based on the TaqMan chemistry was developed for reliable detection and quantitation of Citrus leaf blotch virus (CLBV) in citrus plants. Detection by this method was highly specific and about one thousand times more sensitive than detection by conventional RT-PCR. An external standard curve using in vitro synthesized RNA transcripts of the selected target allowed a reproducible quantitative assay, with a wide dynamic range (seven logarithmic units of concentration) and very low variation coefficient values. This protocol enabled detection of as little as 100 copies of CLBV RNA in various tissues and citrus varieties infected with CLBV sources from different geographical origins. The new assay greatly improves current detection methods for CLBV and it has been most helpful for the Spanish citrus sanitation, quarantine and certification programs, and fitness evaluation of infectious cDNA clones of CLBV, useful potentially as viral vectors for citrus. (C) 2009 Elsevier B.V. All rights reserved. |
| format | Artículo |
| id | ReDivia4456 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia44562025-04-25T14:43:35Z Detection and quantitation of Citrus leaf blotch virus by TaqMan real-time RT-PCR Ruiz-Ruiz, Susana Ambrós, Silvia Vives, María C. Navarro, Luis Moreno, Pedro Guerri, José A real-time RT-PCR assay based on the TaqMan chemistry was developed for reliable detection and quantitation of Citrus leaf blotch virus (CLBV) in citrus plants. Detection by this method was highly specific and about one thousand times more sensitive than detection by conventional RT-PCR. An external standard curve using in vitro synthesized RNA transcripts of the selected target allowed a reproducible quantitative assay, with a wide dynamic range (seven logarithmic units of concentration) and very low variation coefficient values. This protocol enabled detection of as little as 100 copies of CLBV RNA in various tissues and citrus varieties infected with CLBV sources from different geographical origins. The new assay greatly improves current detection methods for CLBV and it has been most helpful for the Spanish citrus sanitation, quarantine and certification programs, and fitness evaluation of infectious cDNA clones of CLBV, useful potentially as viral vectors for citrus. (C) 2009 Elsevier B.V. All rights reserved. 2017-06-01T10:10:10Z 2017-06-01T10:10:10Z 2009 SEP article Ruiz-Ruiz, S., Ambros, S., Vives, M.C., Navarro, L., Moreno, P., Guerri, J. (2009). Detection and quantitation of Citrus leaf blotch virus by TaqMan real-time RT-PCR. Journal of virological methods, 160(1-2), 57-62. 0166-0934 http://hdl.handle.net/20.500.11939/4456 10.1016/j.jviromet.2009.04.012 en openAccess Impreso |
| spellingShingle | Ruiz-Ruiz, Susana Ambrós, Silvia Vives, María C. Navarro, Luis Moreno, Pedro Guerri, José Detection and quantitation of Citrus leaf blotch virus by TaqMan real-time RT-PCR |
| title | Detection and quantitation of Citrus leaf blotch virus by TaqMan real-time RT-PCR |
| title_full | Detection and quantitation of Citrus leaf blotch virus by TaqMan real-time RT-PCR |
| title_fullStr | Detection and quantitation of Citrus leaf blotch virus by TaqMan real-time RT-PCR |
| title_full_unstemmed | Detection and quantitation of Citrus leaf blotch virus by TaqMan real-time RT-PCR |
| title_short | Detection and quantitation of Citrus leaf blotch virus by TaqMan real-time RT-PCR |
| title_sort | detection and quantitation of citrus leaf blotch virus by taqman real time rt pcr |
| url | http://hdl.handle.net/20.500.11939/4456 |
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