Direct sample preparation methods for the detection of Plum pox virus by real-time RT-PCR
Direct systems to process plant materials allowed high-throughput testing of Plum pox virus (PPV) by real-time reverse transcription (RT)-PCR without nucleic acids purification. Crude plant extracts were diluted in buffer or spotted on membranes to be used as templates. Alternatively, immobilized PP...
| Autores principales: | , , , , , |
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| Formato: | Artículo |
| Lenguaje: | Inglés |
| Publicado: |
2017
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| Acceso en línea: | http://hdl.handle.net/20.500.11939/4962 |
| _version_ | 1855491842224685056 |
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| author | Capote, Nieves Bertolini, Edson Olmos, Antonio Vidal, Eduardo Martínez, M. Carmen Cambra, Mariano |
| author_browse | Bertolini, Edson Cambra, Mariano Capote, Nieves Martínez, M. Carmen Olmos, Antonio Vidal, Eduardo |
| author_facet | Capote, Nieves Bertolini, Edson Olmos, Antonio Vidal, Eduardo Martínez, M. Carmen Cambra, Mariano |
| author_sort | Capote, Nieves |
| collection | ReDivia |
| description | Direct systems to process plant materials allowed high-throughput testing of Plum pox virus (PPV) by real-time reverse transcription (RT)-PCR without nucleic acids purification. Crude plant extracts were diluted in buffer or spotted on membranes to be used as templates. Alternatively, immobilized PPV targets were amplified from fresh sections of plant tissues printed or squashed onto the same supports, without extract preparation. Spot real-time RT-PCR was validated as a PPV diagnostic method in samples collected during the dormancy period and showed high sensitivity (93.6%), specificity (98.0%), and post-test probability (97.9%) towards sharka disease. In an analysis of 2919 Prunus samples by spot real-time RT-PCR and DASI-ELISA 90.8% of the results coincided, demonstrating high agreement (k = 0.77 +/- 0.01) between the two techniques. These results validate the use of immobilized PPV targets and spot real-time RT-PCR as screening method for large-scale analyses. [Int Microbiol 2009; 12(1): 1-6] |
| format | Artículo |
| id | ReDivia4962 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia49622025-04-25T14:45:01Z Direct sample preparation methods for the detection of Plum pox virus by real-time RT-PCR Capote, Nieves Bertolini, Edson Olmos, Antonio Vidal, Eduardo Martínez, M. Carmen Cambra, Mariano Direct systems to process plant materials allowed high-throughput testing of Plum pox virus (PPV) by real-time reverse transcription (RT)-PCR without nucleic acids purification. Crude plant extracts were diluted in buffer or spotted on membranes to be used as templates. Alternatively, immobilized PPV targets were amplified from fresh sections of plant tissues printed or squashed onto the same supports, without extract preparation. Spot real-time RT-PCR was validated as a PPV diagnostic method in samples collected during the dormancy period and showed high sensitivity (93.6%), specificity (98.0%), and post-test probability (97.9%) towards sharka disease. In an analysis of 2919 Prunus samples by spot real-time RT-PCR and DASI-ELISA 90.8% of the results coincided, demonstrating high agreement (k = 0.77 +/- 0.01) between the two techniques. These results validate the use of immobilized PPV targets and spot real-time RT-PCR as screening method for large-scale analyses. [Int Microbiol 2009; 12(1): 1-6] 2017-06-01T10:11:27Z 2017-06-01T10:11:27Z 2009 MAR 2009 article publishedVersion Capote, N., Bertolini, E., Olmos, A., Vidal, E., Martinez, M.C., Cambra, M. (2009). Direct sample preparation methods for the detection of Plum pox virus by real-time RT-PCR. International Microbiology, 12(1), 1-6. 1139-6709 http://hdl.handle.net/20.500.11939/4962 http://hdl.handle.net/20.500.11939/4962 10.2436/20.1501.01.75 en openAccess Impreso |
| spellingShingle | Capote, Nieves Bertolini, Edson Olmos, Antonio Vidal, Eduardo Martínez, M. Carmen Cambra, Mariano Direct sample preparation methods for the detection of Plum pox virus by real-time RT-PCR |
| title | Direct sample preparation methods for the detection of Plum pox virus by real-time RT-PCR |
| title_full | Direct sample preparation methods for the detection of Plum pox virus by real-time RT-PCR |
| title_fullStr | Direct sample preparation methods for the detection of Plum pox virus by real-time RT-PCR |
| title_full_unstemmed | Direct sample preparation methods for the detection of Plum pox virus by real-time RT-PCR |
| title_short | Direct sample preparation methods for the detection of Plum pox virus by real-time RT-PCR |
| title_sort | direct sample preparation methods for the detection of plum pox virus by real time rt pcr |
| url | http://hdl.handle.net/20.500.11939/4962 http://hdl.handle.net/20.500.11939/4962 |
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