Simultaneous detection and typing of plum pox potyvirus (PPV) isolates by heminested-PCR and PCR-ELISA
Two techniques for simultaneous detection and typing of plum pox potyvirus (PPV) isolates belonging to the D or M serotypes, heminested PCR (H-PCR) and PCR-ELISA, have been developed. Ten PPV isolates typed using PPV-D and PPV-M specific monoclonal antibodies by ELISA-DASI were used to validate thes...
| Main Authors: | , , , , , , |
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| Format: | article |
| Language: | Inglés |
| Published: |
2017
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| Online Access: | http://hdl.handle.net/20.500.11939/4259 |
| _version_ | 1855032090679050240 |
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| author | Olmos, Antonio Cambra, Mariano Dasi, M. A. Candresse, Thierry Esteban, Olga Gorris, María T. Asensio, M. |
| author_browse | Asensio, M. Cambra, Mariano Candresse, Thierry Dasi, M. A. Esteban, Olga Gorris, María T. Olmos, Antonio |
| author_facet | Olmos, Antonio Cambra, Mariano Dasi, M. A. Candresse, Thierry Esteban, Olga Gorris, María T. Asensio, M. |
| author_sort | Olmos, Antonio |
| collection | ReDivia |
| description | Two techniques for simultaneous detection and typing of plum pox potyvirus (PPV) isolates belonging to the D or M serotypes, heminested PCR (H-PCR) and PCR-ELISA, have been developed. Ten PPV isolates typed using PPV-D and PPV-M specific monoclonal antibodies by ELISA-DASI were used to validate these two methods. The results obtained show a complete coincidence of the nucleic acid-based techniques with the serological data. When serial dilutions of infected plant extracts were assayed, H-PCR and PCR-ELISA were found to be 100 rimes more sensitive than the more conventional immunocapture-PCR (IC-PCR) assay. Testing of 228 PPV-infected fruit tree samples coming from different hosts and locations indicated that so fat-only PPV type D appears to be present in Spain and in Chile. Coupled with print-capture sample preparation (Olmos et al., Nucl. Acids Res. 24, 2192-2193, 1996) the increased sensitivity provided by heminested-PCR allowed the detection of PPV targets of D and M types, in wingless individuals of the aphid vector Aphis gossypii. (C) 1997 Elsevier Science Ireland Ltd. |
| format | article |
| id | ReDivia4259 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia42592025-04-25T14:42:40Z Simultaneous detection and typing of plum pox potyvirus (PPV) isolates by heminested-PCR and PCR-ELISA Olmos, Antonio Cambra, Mariano Dasi, M. A. Candresse, Thierry Esteban, Olga Gorris, María T. Asensio, M. Two techniques for simultaneous detection and typing of plum pox potyvirus (PPV) isolates belonging to the D or M serotypes, heminested PCR (H-PCR) and PCR-ELISA, have been developed. Ten PPV isolates typed using PPV-D and PPV-M specific monoclonal antibodies by ELISA-DASI were used to validate these two methods. The results obtained show a complete coincidence of the nucleic acid-based techniques with the serological data. When serial dilutions of infected plant extracts were assayed, H-PCR and PCR-ELISA were found to be 100 rimes more sensitive than the more conventional immunocapture-PCR (IC-PCR) assay. Testing of 228 PPV-infected fruit tree samples coming from different hosts and locations indicated that so fat-only PPV type D appears to be present in Spain and in Chile. Coupled with print-capture sample preparation (Olmos et al., Nucl. Acids Res. 24, 2192-2193, 1996) the increased sensitivity provided by heminested-PCR allowed the detection of PPV targets of D and M types, in wingless individuals of the aphid vector Aphis gossypii. (C) 1997 Elsevier Science Ireland Ltd. 2017-06-01T10:09:42Z 2017-06-01T10:09:42Z 1997 NOV 1997 article Olmos, A., Cambra, M., Dasi, M.A., Candresse, T., Esteban, O., Gorris, M. T., Asensio, M. (1997). Simultaneous detection and typing of plum pox potyvirus (PPV), isolates by heminested-PCR and PCR-ELISA. Journal of virological methods, 68(2), 127-137. 0166-0934 http://hdl.handle.net/20.500.11939/4259 10.1016/S0166-0934(97)00120-1 en openAccess Impreso |
| spellingShingle | Olmos, Antonio Cambra, Mariano Dasi, M. A. Candresse, Thierry Esteban, Olga Gorris, María T. Asensio, M. Simultaneous detection and typing of plum pox potyvirus (PPV) isolates by heminested-PCR and PCR-ELISA |
| title | Simultaneous detection and typing of plum pox potyvirus (PPV) isolates by heminested-PCR and PCR-ELISA |
| title_full | Simultaneous detection and typing of plum pox potyvirus (PPV) isolates by heminested-PCR and PCR-ELISA |
| title_fullStr | Simultaneous detection and typing of plum pox potyvirus (PPV) isolates by heminested-PCR and PCR-ELISA |
| title_full_unstemmed | Simultaneous detection and typing of plum pox potyvirus (PPV) isolates by heminested-PCR and PCR-ELISA |
| title_short | Simultaneous detection and typing of plum pox potyvirus (PPV) isolates by heminested-PCR and PCR-ELISA |
| title_sort | simultaneous detection and typing of plum pox potyvirus ppv isolates by heminested pcr and pcr elisa |
| url | http://hdl.handle.net/20.500.11939/4259 |
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