Quantitative detection of Citrus tristeza virus in plant tissues and single aphids by real-time RT-PCR
TaqMan real-time reverse transcriptase (RT)-polymerase chain reaction (PCR) using purified RNA targets or coupled to tissue-print and squash procedures was developed to detect and quantify Citrus tristeza virus (CTV) RNA-targets in plant tissues and in single aphids. With this method all CTV isolate...
| Autores principales: | , , , , , , , |
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| Formato: | article |
| Lenguaje: | Inglés |
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2017
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| Acceso en línea: | http://hdl.handle.net/20.500.11939/4841 |
| _version_ | 1855032185591955456 |
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| author | Bertolini, Edson Moreno, Aránzazu Capote, Nieves Olmos, Antonio de Luis, Ana Vidal, Eduardo Pérez-Panadés, J. Cambra, Mariano |
| author_browse | Bertolini, Edson Cambra, Mariano Capote, Nieves Moreno, Aránzazu Olmos, Antonio Pérez-Panadés, J. Vidal, Eduardo de Luis, Ana |
| author_facet | Bertolini, Edson Moreno, Aránzazu Capote, Nieves Olmos, Antonio de Luis, Ana Vidal, Eduardo Pérez-Panadés, J. Cambra, Mariano |
| author_sort | Bertolini, Edson |
| collection | ReDivia |
| description | TaqMan real-time reverse transcriptase (RT)-polymerase chain reaction (PCR) using purified RNA targets or coupled to tissue-print and squash procedures was developed to detect and quantify Citrus tristeza virus (CTV) RNA-targets in plant tissues and in single aphids. With this method all CTV isolates tested from different hosts and origins were detected. The sensitivity of conventional real-time RT-PCR was 1,000 times higher than immunocapture (IC)-RT-nested PCR and 10(6) times higher than enzyme linked immunosorbent assay (ELISA). The quantitation limit ranged from 1.7x10(2) to 1.7x10(9) transcript copies. The estimated number of CTV RNA-targets detected in different organs of a CTV-infected tree ranged from 4.5x10(5) to 6.5x10(8) copies when purified RNA was used as template and from 1.9x10(4) to 3.7x10(6) when tissue-printed material was used. In single squashed aphids the number of copies ranged from 4.73x10(3) to 1.23x10(5). Reliable quantitation of CTV targets present in infected plant material or acquired by single aphid species, was achieved with tissue-print and squash procedures combined with real-time RT-PCR, both of which do not require extraction procedures or nucleic acid purification. |
| format | article |
| id | ReDivia4841 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia48412025-04-25T14:44:45Z Quantitative detection of Citrus tristeza virus in plant tissues and single aphids by real-time RT-PCR Bertolini, Edson Moreno, Aránzazu Capote, Nieves Olmos, Antonio de Luis, Ana Vidal, Eduardo Pérez-Panadés, J. Cambra, Mariano TaqMan real-time reverse transcriptase (RT)-polymerase chain reaction (PCR) using purified RNA targets or coupled to tissue-print and squash procedures was developed to detect and quantify Citrus tristeza virus (CTV) RNA-targets in plant tissues and in single aphids. With this method all CTV isolates tested from different hosts and origins were detected. The sensitivity of conventional real-time RT-PCR was 1,000 times higher than immunocapture (IC)-RT-nested PCR and 10(6) times higher than enzyme linked immunosorbent assay (ELISA). The quantitation limit ranged from 1.7x10(2) to 1.7x10(9) transcript copies. The estimated number of CTV RNA-targets detected in different organs of a CTV-infected tree ranged from 4.5x10(5) to 6.5x10(8) copies when purified RNA was used as template and from 1.9x10(4) to 3.7x10(6) when tissue-printed material was used. In single squashed aphids the number of copies ranged from 4.73x10(3) to 1.23x10(5). Reliable quantitation of CTV targets present in infected plant material or acquired by single aphid species, was achieved with tissue-print and squash procedures combined with real-time RT-PCR, both of which do not require extraction procedures or nucleic acid purification. 2017-06-01T10:11:09Z 2017-06-01T10:11:09Z 2008 FEB 2008 article Bertolini, E., Moreno, A., Capote, N., Olmos, A., de L., Ana, Vidal, E., Perez-Panades, Jordi, Cambra, M. (2008). Quantitative detection of Citrus tristeza virus in plant tissues and single aphids by real-time RT-PCR. European Journal of Plant Pathology, 120(2), 177-188. 0929-1873 http://hdl.handle.net/20.500.11939/4841 10.1007/s10658-007-9206-9 en openAccess Impreso |
| spellingShingle | Bertolini, Edson Moreno, Aránzazu Capote, Nieves Olmos, Antonio de Luis, Ana Vidal, Eduardo Pérez-Panadés, J. Cambra, Mariano Quantitative detection of Citrus tristeza virus in plant tissues and single aphids by real-time RT-PCR |
| title | Quantitative detection of Citrus tristeza virus in plant tissues and single aphids by real-time RT-PCR |
| title_full | Quantitative detection of Citrus tristeza virus in plant tissues and single aphids by real-time RT-PCR |
| title_fullStr | Quantitative detection of Citrus tristeza virus in plant tissues and single aphids by real-time RT-PCR |
| title_full_unstemmed | Quantitative detection of Citrus tristeza virus in plant tissues and single aphids by real-time RT-PCR |
| title_short | Quantitative detection of Citrus tristeza virus in plant tissues and single aphids by real-time RT-PCR |
| title_sort | quantitative detection of citrus tristeza virus in plant tissues and single aphids by real time rt pcr |
| url | http://hdl.handle.net/20.500.11939/4841 |
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