Loop-Mediated Isothermal Amplification of Specific Endoglucanase Gene Sequence for Detection of the Bacterial Wilt Pathogen Ralstonia solanacearum
The increased globalization of crops production and processing industries also promotes the side-effects of more rapid and efficient spread of plant pathogens. To prevent the associated economic losses, and particularly those related to bacterial diseases where their management relies on removal of...
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| Format: | article |
| Language: | Inglés |
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PLOS
2022
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| Online Access: | http://hdl.handle.net/20.500.11939/8062 https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0096027 |
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| author | Lenarcic, Rok Morisset, Dany Pirc, Mnaca Llop, Pablo Ravnikar, Maja Dreo, Tanja |
| author_browse | Dreo, Tanja Lenarcic, Rok Llop, Pablo Morisset, Dany Pirc, Mnaca Ravnikar, Maja |
| author_facet | Lenarcic, Rok Morisset, Dany Pirc, Mnaca Llop, Pablo Ravnikar, Maja Dreo, Tanja |
| author_sort | Lenarcic, Rok |
| collection | ReDivia |
| description | The increased globalization of crops production and processing industries also promotes the side-effects of more rapid and efficient spread of plant pathogens. To prevent the associated economic losses, and particularly those related to bacterial diseases where their management relies on removal of the infected material from production, simple, easy-to-perform, rapid and cost-effective tests are needed. Loop-mediated isothermal amplification (LAMP) assays that target 16S rRNA, fliC and egl genes were compared and evaluated as on-site applications. The assay with the best performance was that targeted to the egl gene, which shows high analytical specificity for diverse strains of the betaproteobacterium Ralstonia solanacearum, including its non-European and non-race 3 biovar 2 strains. The additional melting curve analysis provides confirmation of the test results. According to our extensive assessment, the egl LAMP assay requires minimum sample preparation (a few minutes of boiling) for the identification of pure cultures and ooze from symptomatic material, and it can also be used in a high-throughput format in the laboratory. This provides sensitive and reliable detection of R. solanacearum strains of different phylotypes. |
| format | article |
| id | ReDivia8062 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2022 |
| publishDateRange | 2022 |
| publishDateSort | 2022 |
| publisher | PLOS |
| publisherStr | PLOS |
| record_format | dspace |
| spelling | ReDivia80622025-04-25T14:48:44Z Loop-Mediated Isothermal Amplification of Specific Endoglucanase Gene Sequence for Detection of the Bacterial Wilt Pathogen Ralstonia solanacearum Lenarcic, Rok Morisset, Dany Pirc, Mnaca Llop, Pablo Ravnikar, Maja Dreo, Tanja Loop-mediated isothermal amplification U30 Research methods H20 Plant diseases Gene sequence Ralstonia solanacearum Diagnostic techniques PCR Testing The increased globalization of crops production and processing industries also promotes the side-effects of more rapid and efficient spread of plant pathogens. To prevent the associated economic losses, and particularly those related to bacterial diseases where their management relies on removal of the infected material from production, simple, easy-to-perform, rapid and cost-effective tests are needed. Loop-mediated isothermal amplification (LAMP) assays that target 16S rRNA, fliC and egl genes were compared and evaluated as on-site applications. The assay with the best performance was that targeted to the egl gene, which shows high analytical specificity for diverse strains of the betaproteobacterium Ralstonia solanacearum, including its non-European and non-race 3 biovar 2 strains. The additional melting curve analysis provides confirmation of the test results. According to our extensive assessment, the egl LAMP assay requires minimum sample preparation (a few minutes of boiling) for the identification of pure cultures and ooze from symptomatic material, and it can also be used in a high-throughput format in the laboratory. This provides sensitive and reliable detection of R. solanacearum strains of different phylotypes. 2022-04-29T16:21:20Z 2022-04-29T16:21:20Z 2014 article publishedVersion Lenarčič, R., Morisset, D., Pirc, M., Llop, P., Ravnikar, M. & Dreo, T. (2014). Loop-mediated isothermal amplification of specific endoglucanase gene sequence for detection of the bacterial wilt pathogen Ralstonia solanacearum. PLoS One, 9(4), e96027. 1932-6203 (eISSN) http://hdl.handle.net/20.500.11939/8062 10.1371/journal.pone.0096027 https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0096027 en info:eu-repo/grantAgreement/EC/FP7/245047 This study was carried out under the sponsorship of the EU Framework 7 Programme (FP7-KBBE-2009-3) project 245047 (Qdetect – Developing quarantine pest detection methods for use by national plant protection organisations – NPPO and inspection services) and of the Slovenian Research Agency funded project Q-finder L1-3642 (Developing simple, rapid and on-site methods for plant pathogens detection). Atribución-NoComercial-SinDerivadas 3.0 España http://creativecommons.org/licenses/by-nc-nd/3.0/es/ openAccess PLOS electronico |
| spellingShingle | Loop-mediated isothermal amplification U30 Research methods H20 Plant diseases Gene sequence Ralstonia solanacearum Diagnostic techniques PCR Testing Lenarcic, Rok Morisset, Dany Pirc, Mnaca Llop, Pablo Ravnikar, Maja Dreo, Tanja Loop-Mediated Isothermal Amplification of Specific Endoglucanase Gene Sequence for Detection of the Bacterial Wilt Pathogen Ralstonia solanacearum |
| title | Loop-Mediated Isothermal Amplification of Specific Endoglucanase Gene Sequence for Detection of the Bacterial Wilt Pathogen Ralstonia solanacearum |
| title_full | Loop-Mediated Isothermal Amplification of Specific Endoglucanase Gene Sequence for Detection of the Bacterial Wilt Pathogen Ralstonia solanacearum |
| title_fullStr | Loop-Mediated Isothermal Amplification of Specific Endoglucanase Gene Sequence for Detection of the Bacterial Wilt Pathogen Ralstonia solanacearum |
| title_full_unstemmed | Loop-Mediated Isothermal Amplification of Specific Endoglucanase Gene Sequence for Detection of the Bacterial Wilt Pathogen Ralstonia solanacearum |
| title_short | Loop-Mediated Isothermal Amplification of Specific Endoglucanase Gene Sequence for Detection of the Bacterial Wilt Pathogen Ralstonia solanacearum |
| title_sort | loop mediated isothermal amplification of specific endoglucanase gene sequence for detection of the bacterial wilt pathogen ralstonia solanacearum |
| topic | Loop-mediated isothermal amplification U30 Research methods H20 Plant diseases Gene sequence Ralstonia solanacearum Diagnostic techniques PCR Testing |
| url | http://hdl.handle.net/20.500.11939/8062 https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0096027 |
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