A new and sensitive co-operational polymerase chain reaction for rapid detection of Ralstonia solanacearum in water
Three primers from 16S rRNA were successfully assayed simultaneously in one reaction for sensitive detection of Ralstonia solanacearum in watercourses. The protocol is a modification of the Co-operational polymerase chain reaction (Co-PCR), which allows the simultaneous and co-operational action of...
| Autores principales: | , , , |
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| Formato: | article |
| Lenguaje: | Inglés |
| Publicado: |
2017
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| Acceso en línea: | http://hdl.handle.net/20.500.11939/4976 https://www.sciencedirect.com/science/article/abs/pii/S0167701203001611?via%3Dihub |
| _version_ | 1855032205494976512 |
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| author | Caruso, Paola Bertolini, Edson Cambra, Mariano López, María M. |
| author_browse | Bertolini, Edson Cambra, Mariano Caruso, Paola López, María M. |
| author_facet | Caruso, Paola Bertolini, Edson Cambra, Mariano López, María M. |
| author_sort | Caruso, Paola |
| collection | ReDivia |
| description | Three primers from 16S rRNA were successfully assayed simultaneously in one reaction for sensitive detection of Ralstonia solanacearum in watercourses. The protocol is a modification of the Co-operational polymerase chain reaction (Co-PCR), which allows the simultaneous and co-operational action of the primers. It specifically amplified R. solanacearum strains belonging to biovars 1, 2 and 4. No products were obtained from any of the 162 unidentified isolates from river water. The sensitivity of the assay was <1 cfu/ml as determined by analysis of heat-treated water samples spiked with R. solanacearum, also containing indigenous microbiota up to 105 cfu/ml. The developed Co-PCR assay was more sensitive than other standard PCR assays in the analysis of 51 Spanish environmental water samples. Namely 31.3% of the samples were positive using the newly developed assay, whereas 13.7% or less positive samples were found with the other protocols. The Co-PCR improves the detection sensitivity of R. solanacearum and provides an important tool for its routine detection from environmental water samples and for epidemiological studies. |
| format | article |
| id | ReDivia4976 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | ReDivia49762025-04-25T14:45:04Z A new and sensitive co-operational polymerase chain reaction for rapid detection of Ralstonia solanacearum in water Caruso, Paola Bertolini, Edson Cambra, Mariano López, María M. Three primers from 16S rRNA were successfully assayed simultaneously in one reaction for sensitive detection of Ralstonia solanacearum in watercourses. The protocol is a modification of the Co-operational polymerase chain reaction (Co-PCR), which allows the simultaneous and co-operational action of the primers. It specifically amplified R. solanacearum strains belonging to biovars 1, 2 and 4. No products were obtained from any of the 162 unidentified isolates from river water. The sensitivity of the assay was <1 cfu/ml as determined by analysis of heat-treated water samples spiked with R. solanacearum, also containing indigenous microbiota up to 105 cfu/ml. The developed Co-PCR assay was more sensitive than other standard PCR assays in the analysis of 51 Spanish environmental water samples. Namely 31.3% of the samples were positive using the newly developed assay, whereas 13.7% or less positive samples were found with the other protocols. The Co-PCR improves the detection sensitivity of R. solanacearum and provides an important tool for its routine detection from environmental water samples and for epidemiological studies. 2017-06-01T10:11:29Z 2017-06-01T10:11:29Z 2003 OCT 2003 article Caruso, P., Bertolini, E., Cambra, M., Lopez, M.M. (2003). A new and sensitive co-operational polymerase chain reaction for rapid detection of Ralstonia solanacearum in water. Journal of microbiological methods, 55(1), 257-272. 0167-7012 http://hdl.handle.net/20.500.11939/4976 10.1016/S0167-7012(03)00161-1 https://www.sciencedirect.com/science/article/abs/pii/S0167701203001611?via%3Dihub en openAccess Impreso |
| spellingShingle | Caruso, Paola Bertolini, Edson Cambra, Mariano López, María M. A new and sensitive co-operational polymerase chain reaction for rapid detection of Ralstonia solanacearum in water |
| title | A new and sensitive co-operational polymerase chain reaction for rapid detection of Ralstonia solanacearum in water |
| title_full | A new and sensitive co-operational polymerase chain reaction for rapid detection of Ralstonia solanacearum in water |
| title_fullStr | A new and sensitive co-operational polymerase chain reaction for rapid detection of Ralstonia solanacearum in water |
| title_full_unstemmed | A new and sensitive co-operational polymerase chain reaction for rapid detection of Ralstonia solanacearum in water |
| title_short | A new and sensitive co-operational polymerase chain reaction for rapid detection of Ralstonia solanacearum in water |
| title_sort | new and sensitive co operational polymerase chain reaction for rapid detection of ralstonia solanacearum in water |
| url | http://hdl.handle.net/20.500.11939/4976 https://www.sciencedirect.com/science/article/abs/pii/S0167701203001611?via%3Dihub |
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