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
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/8234
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oai_identifier_str oai:localhost:20.500.12123/8234
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spelling 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)2025-09-29T13:45:03Zoai: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:l2025-09-29 13:45:04.318INTA 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
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str 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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score 12.558318

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