Isothermal amplification coupled with rapid flow-through hybridisation for sensitive diagnosis of Plum pox virus
A nucleic acid sequence-based amplification method Coupled with rapid flow-through hybridisation (NASBA-FH) was developed for diagnosis of Plum pox virus (PPV). The sensitivity level achieved by NASBA-FH was 10 times higher than that obtained by Co-PCR and 1000 times higher than the sensitivity affo...
| Autores principales: | , , |
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| Formato: | Artículo |
| Lenguaje: | Inglés |
| Publicado: |
2017
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| Acceso en línea: | http://hdl.handle.net/20.500.11939/4258 |
| _version_ | 1855491721397272576 |
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| author | Olmos, Antonio Bertolini, Edson Cambra, Mariano |
| author_browse | Bertolini, Edson Cambra, Mariano Olmos, Antonio |
| author_facet | Olmos, Antonio Bertolini, Edson Cambra, Mariano |
| author_sort | Olmos, Antonio |
| collection | ReDivia |
| description | A nucleic acid sequence-based amplification method Coupled with rapid flow-through hybridisation (NASBA-FH) was developed for diagnosis of Plum pox virus (PPV). The sensitivity level achieved by NASBA-FH was 10 times higher than that obtained by Co-PCR and 1000 times higher than the sensitivity afforded by RT-PCR. In addition, samples from 262 stone-fruit trees collected during winter and spring seasons were analysed. These samples were tested using methods recommended by the European and Mediterranean Plant Protection Organization to detect PPV (DASI-ELISA, RT-PCR and Co-PCR) and by NASBA-FH. Winter PPV diagnostic results by ELISA and NASBA-FH coincided in 90.8%, while ELISA and PCR-based methods coincided in 91.6% and PCR-based methods with NASBA-FH agreed in 95.4%. In spring, diagnostic results were similar with all the molecular techniques, which agreed with ELISA results for 98.8% of the trees. NASBA-FH was able to detect more positive infections in winter, which were later confirmed in spring. These results indicate that NASBA-FH is a suitable molecular method for routine PPV detection in the winter and spring. This user-friendly isothermal RNA amplification coupled with a very fast flow-through hybridisation (15 min) opens up new possibilities for rapid and reliable diagnosis of a variety of pathogens. (c) 2006 Elsevier B.V. All rights reserved. |
| format | Artículo |
| id | ReDivia4258 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia42582025-04-25T14:42:40Z Isothermal amplification coupled with rapid flow-through hybridisation for sensitive diagnosis of Plum pox virus Olmos, Antonio Bertolini, Edson Cambra, Mariano A nucleic acid sequence-based amplification method Coupled with rapid flow-through hybridisation (NASBA-FH) was developed for diagnosis of Plum pox virus (PPV). The sensitivity level achieved by NASBA-FH was 10 times higher than that obtained by Co-PCR and 1000 times higher than the sensitivity afforded by RT-PCR. In addition, samples from 262 stone-fruit trees collected during winter and spring seasons were analysed. These samples were tested using methods recommended by the European and Mediterranean Plant Protection Organization to detect PPV (DASI-ELISA, RT-PCR and Co-PCR) and by NASBA-FH. Winter PPV diagnostic results by ELISA and NASBA-FH coincided in 90.8%, while ELISA and PCR-based methods coincided in 91.6% and PCR-based methods with NASBA-FH agreed in 95.4%. In spring, diagnostic results were similar with all the molecular techniques, which agreed with ELISA results for 98.8% of the trees. NASBA-FH was able to detect more positive infections in winter, which were later confirmed in spring. These results indicate that NASBA-FH is a suitable molecular method for routine PPV detection in the winter and spring. This user-friendly isothermal RNA amplification coupled with a very fast flow-through hybridisation (15 min) opens up new possibilities for rapid and reliable diagnosis of a variety of pathogens. (c) 2006 Elsevier B.V. All rights reserved. 2017-06-01T10:09:42Z 2017-06-01T10:09:42Z 2007 JAN 2007 article Olmos, A., Bertolini, E., Cambra, M. (2007). Isothermal amplification coupled with rapid flow-through hybridisation for sensitive diagnosis of Plum pox virus. Journal of virological methods, 139(1), 111-115. 0166-0934 http://hdl.handle.net/20.500.11939/4258 10.1016/j.jviromet.2006.09.012 en openAccess Impreso |
| spellingShingle | Olmos, Antonio Bertolini, Edson Cambra, Mariano Isothermal amplification coupled with rapid flow-through hybridisation for sensitive diagnosis of Plum pox virus |
| title | Isothermal amplification coupled with rapid flow-through hybridisation for sensitive diagnosis of Plum pox virus |
| title_full | Isothermal amplification coupled with rapid flow-through hybridisation for sensitive diagnosis of Plum pox virus |
| title_fullStr | Isothermal amplification coupled with rapid flow-through hybridisation for sensitive diagnosis of Plum pox virus |
| title_full_unstemmed | Isothermal amplification coupled with rapid flow-through hybridisation for sensitive diagnosis of Plum pox virus |
| title_short | Isothermal amplification coupled with rapid flow-through hybridisation for sensitive diagnosis of Plum pox virus |
| title_sort | isothermal amplification coupled with rapid flow through hybridisation for sensitive diagnosis of plum pox virus |
| url | http://hdl.handle.net/20.500.11939/4258 |
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