LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis
In this study, we developed new sets of primers to detect Brucella spp. and M. avium subsp. paratuberculosis (MAP) through isothermal amplification. We selected a previously well-characterized target gene, bscp31, specific for Brucella spp. and IS900 for MAP. The limits of detection using the loop-m...
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| Format: | Artículo |
| Language: | Inglés |
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Sociedade Brasileira de Microbiologia
2020
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| Online Access: | https://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822015000200619&lng=en&nrm=iso&tlng=en http://hdl.handle.net/20.500.12123/8234 https://doi.org/10.1590/S1517-838246220131206 |
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| author | Trangoni, Marcos David Gioffre, Andrea Ceron Cucchi, Maria Esperanza Caimi, Karina Cynthia Ruybal, Paula Zumarraga, Martin Jose Cravero, Silvio Lorenzo Pedro |
| author_browse | Caimi, Karina Cynthia Ceron Cucchi, Maria Esperanza Cravero, Silvio Lorenzo Pedro Gioffre, Andrea Ruybal, Paula Trangoni, Marcos David Zumarraga, Martin Jose |
| author_facet | Trangoni, Marcos David Gioffre, Andrea Ceron Cucchi, Maria Esperanza Caimi, Karina Cynthia Ruybal, Paula Zumarraga, Martin Jose Cravero, Silvio Lorenzo Pedro |
| author_sort | Trangoni, Marcos David |
| collection | INTA Digital |
| description | In this study, we developed new sets of primers to detect Brucella spp. and M. avium subsp. paratuberculosis (MAP) through isothermal amplification. We selected a previously well-characterized target gene, bscp31, specific for Brucella spp. and IS900 for MAP. The limits of detection using the loop-mediated isothermal amplification (LAMP) protocols described herein were similar to those of conventional PCR targeting the same sequences. Hydroxynaphtol blue and SYBR GreenTM allowed direct naked-eye detection with identical sensitivity as agarose gel electrophoresis. We included the LAMP-based protocol in a rapid identification scheme of the respective pathogens, and all tested isolates were correctly identified within 2 to 3 h. In addition, both protocols were suitable for specifically identifying the respective pathogens; in the case of Brucella, it also allowed the identification of all the biovars tested. We conclude that LAMP is a suitable rapid molecular typing tool that could help to shorten the time required to identify insidious bacteria in low-complexity laboratories, mainly in developing countries. |
| format | Artículo |
| id | INTA8234 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2020 |
| publishDateRange | 2020 |
| publishDateSort | 2020 |
| publisher | Sociedade Brasileira de Microbiologia |
| publisherStr | Sociedade Brasileira de Microbiologia |
| record_format | dspace |
| spelling | INTA82342020-11-10T16:41:32Z LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis Trangoni, Marcos David Gioffre, Andrea Ceron Cucchi, Maria Esperanza Caimi, Karina Cynthia Ruybal, Paula Zumarraga, Martin Jose Cravero, Silvio Lorenzo Pedro Isothermal Processes Molecular Biology Brucellosis Procesos Isotermales Biología Molecular Brucelosis Paratuberculosis Mycobacterium avium subsp. paratuberculosis In this study, we developed new sets of primers to detect Brucella spp. and M. avium subsp. paratuberculosis (MAP) through isothermal amplification. We selected a previously well-characterized target gene, bscp31, specific for Brucella spp. and IS900 for MAP. The limits of detection using the loop-mediated isothermal amplification (LAMP) protocols described herein were similar to those of conventional PCR targeting the same sequences. Hydroxynaphtol blue and SYBR GreenTM allowed direct naked-eye detection with identical sensitivity as agarose gel electrophoresis. We included the LAMP-based protocol in a rapid identification scheme of the respective pathogens, and all tested isolates were correctly identified within 2 to 3 h. In addition, both protocols were suitable for specifically identifying the respective pathogens; in the case of Brucella, it also allowed the identification of all the biovars tested. We conclude that LAMP is a suitable rapid molecular typing tool that could help to shorten the time required to identify insidious bacteria in low-complexity laboratories, mainly in developing countries. Instituto de Biotecnología Fil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Gioffre, Andrea Karina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Ceron Cucchi, Maria Esperanza. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina Fil: Caimi, Karina Cynthia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Ruybal, Paula. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Zumarraga, Martin Jose. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Cravero, Silvio Lorenzo Pedro. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina 2020-11-10T16:35:00Z 2020-11-10T16:35:00Z 2015-06 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion https://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822015000200619&lng=en&nrm=iso&tlng=en http://hdl.handle.net/20.500.12123/8234 1678-4405 https://doi.org/10.1590/S1517-838246220131206 eng info:eu-repograntAgreement/INTA/AERG-232121/AR./Desarrollo de métodos de diagnóstico molecular aplicados a la detección y tipificación de microorganismos patógenos info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Sociedade Brasileira de Microbiologia Brazilian Journal of Microbiology 46 (2) : 619-626 (Junio 2015) |
| spellingShingle | Isothermal Processes Molecular Biology Brucellosis Procesos Isotermales Biología Molecular Brucelosis Paratuberculosis Mycobacterium avium subsp. paratuberculosis Trangoni, Marcos David Gioffre, Andrea Ceron Cucchi, Maria Esperanza Caimi, Karina Cynthia Ruybal, Paula Zumarraga, Martin Jose Cravero, Silvio Lorenzo Pedro LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis |
| title | LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis |
| title_full | LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis |
| title_fullStr | LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis |
| title_full_unstemmed | LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis |
| title_short | LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis |
| title_sort | lamp technology rapid identification of brucella and mycobacterium avium subsp paratuberculosis |
| topic | Isothermal Processes Molecular Biology Brucellosis Procesos Isotermales Biología Molecular Brucelosis Paratuberculosis Mycobacterium avium subsp. paratuberculosis |
| url | https://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822015000200619&lng=en&nrm=iso&tlng=en http://hdl.handle.net/20.500.12123/8234 https://doi.org/10.1590/S1517-838246220131206 |
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