LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis
- Autores
- Trangoni, Marcos David; Gioffre, Andrea; Ceron Cucchi, Maria Esperanza; Caimi, Karina Cynthia; Ruybal, Paula; Zumarraga, Martin Jose; Cravero, Silvio Lorenzo Pedro
- Año de publicación
- 2015
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- 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 - Fuente
- Brazilian Journal of Microbiology 46 (2) : 619-626 (Junio 2015)
- Materia
-
Isothermal Processes
Molecular Biology
Brucellosis
Procesos Isotermales
Biología Molecular
Brucelosis
Paratuberculosis
Mycobacterium avium subsp. paratuberculosis - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- http://creativecommons.org/licenses/by-nc-sa/4.0/
- Repositorio
- Institución
- Instituto Nacional de Tecnología Agropecuaria
- OAI Identificador
- oai:localhost:20.500.12123/8234
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LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosisTrangoni, Marcos DavidGioffre, AndreaCeron Cucchi, Maria EsperanzaCaimi, Karina CynthiaRuybal, PaulaZumarraga, Martin JoseCravero, Silvio Lorenzo PedroIsothermal ProcessesMolecular BiologyBrucellosisProcesos IsotermalesBiología MolecularBrucelosisParatuberculosisMycobacterium avium subsp. paratuberculosisIn 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 BiotecnologíaFil: Trangoni, Marcos David. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Gioffre, Andrea Karina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Ceron Cucchi, Maria Esperanza. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Caimi, Karina Cynthia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Ruybal, Paula. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Zumarraga, Martin Jose. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Cravero, Silvio Lorenzo Pedro. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaSociedade Brasileira de Microbiologia2020-11-10T16:35:00Z2020-11-10T16:35:00Z2015-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttps://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822015000200619&lng=en&nrm=iso&tlng=enhttp://hdl.handle.net/20.500.12123/82341678-4405https://doi.org/10.1590/S1517-838246220131206Brazilian Journal of Microbiology 46 (2) : 619-626 (Junio 2015)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repograntAgreement/INTA/AERG-232121/AR./Desarrollo de métodos de diagnóstico molecular aplicados a la detección y tipificación de microorganismos patógenosinfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2025-09-04T09:48:40Zoai:localhost:20.500.12123/8234instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-04 09:48:42.277INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse |
dc.title.none.fl_str_mv |
LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis |
title |
LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis |
spellingShingle |
LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis Trangoni, Marcos David Isothermal Processes Molecular Biology Brucellosis Procesos Isotermales Biología Molecular Brucelosis Paratuberculosis Mycobacterium avium subsp. paratuberculosis |
title_short |
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_sort |
LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis |
dc.creator.none.fl_str_mv |
Trangoni, Marcos David Gioffre, Andrea Ceron Cucchi, Maria Esperanza Caimi, Karina Cynthia Ruybal, Paula Zumarraga, Martin Jose Cravero, Silvio Lorenzo Pedro |
author |
Trangoni, Marcos David |
author_facet |
Trangoni, Marcos David Gioffre, Andrea Ceron Cucchi, Maria Esperanza Caimi, Karina Cynthia Ruybal, Paula Zumarraga, Martin Jose Cravero, Silvio Lorenzo Pedro |
author_role |
author |
author2 |
Gioffre, Andrea Ceron Cucchi, Maria Esperanza Caimi, Karina Cynthia Ruybal, Paula Zumarraga, Martin Jose Cravero, Silvio Lorenzo Pedro |
author2_role |
author author author author author author |
dc.subject.none.fl_str_mv |
Isothermal Processes Molecular Biology Brucellosis Procesos Isotermales Biología Molecular Brucelosis Paratuberculosis Mycobacterium avium subsp. paratuberculosis |
topic |
Isothermal Processes Molecular Biology Brucellosis Procesos Isotermales Biología Molecular Brucelosis Paratuberculosis Mycobacterium avium subsp. paratuberculosis |
dc.description.none.fl_txt_mv |
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 |
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. |
publishDate |
2015 |
dc.date.none.fl_str_mv |
2015-06 2020-11-10T16:35:00Z 2020-11-10T16:35:00Z |
dc.type.none.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://purl.org/coar/resource_type/c_6501 info:ar-repo/semantics/articulo |
format |
article |
status_str |
publishedVersion |
dc.identifier.none.fl_str_mv |
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 |
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 |
identifier_str_mv |
1678-4405 |
dc.language.none.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
info:eu-repograntAgreement/INTA/AERG-232121/AR./Desarrollo de métodos de diagnóstico molecular aplicados a la detección y tipificación de microorganismos patógenos |
dc.rights.none.fl_str_mv |
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) |
eu_rights_str_mv |
openAccess |
rights_invalid_str_mv |
http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Sociedade Brasileira de Microbiologia |
publisher.none.fl_str_mv |
Sociedade Brasileira de Microbiologia |
dc.source.none.fl_str_mv |
Brazilian Journal of Microbiology 46 (2) : 619-626 (Junio 2015) reponame:INTA Digital (INTA) instname:Instituto Nacional de Tecnología Agropecuaria |
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INTA Digital (INTA) |
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INTA Digital (INTA) |
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Instituto Nacional de Tecnología Agropecuaria |
repository.name.fl_str_mv |
INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria |
repository.mail.fl_str_mv |
tripaldi.nicolas@inta.gob.ar |
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