LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis

Autores
Trangoni, Marcos D.; Gioffré, Andrea Karina; Cerón Cucchi, María Esperanza; Caimi, Karina Cynthia; Ruybal, Paula; Zumárraga, Martín José; 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 Green ™ 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.
Fil: Trangoni, Marcos D.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Gioffré, Andrea Karina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Cerón Cucchi, María Esperanza. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; Argentina
Fil: Caimi, Karina Cynthia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Ruybal, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Zumárraga, Martín José. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Cravero, Silvio Lorenzo Pedro. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Materia
BRUCELLOSIS
LOOP-MEDIATED ISOTHERMAL AMPLIFICATION
MOLECULAR TYPING
PARATUBERCULOSIS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/38587
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/38587
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosisTrangoni, Marcos D.Gioffré, Andrea KarinaCerón Cucchi, María EsperanzaCaimi, Karina CynthiaRuybal, PaulaZumárraga, Martín JoséCravero, Silvio Lorenzo PedroBRUCELLOSISLOOP-MEDIATED ISOTHERMAL AMPLIFICATIONMOLECULAR TYPINGPARATUBERCULOSIShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1In 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 Green ™ 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.Fil: Trangoni, Marcos D.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaFil: Gioffré, Andrea Karina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Cerón Cucchi, María Esperanza. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; ArgentinaFil: Caimi, Karina Cynthia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaFil: Ruybal, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaFil: Zumárraga, Martín José. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaFil: Cravero, Silvio Lorenzo Pedro. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaSociedade Brasileira de Microbiologia2015-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/38587Trangoni, Marcos D.; Gioffré, Andrea Karina; Cerón Cucchi, María Esperanza; Caimi, Karina Cynthia; Ruybal, Paula; et al.; LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis; Sociedade Brasileira de Microbiologia; Brazilian Journal of Microbiology; 46; 2; 6-2015; 619-6261517-83821678-4405CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1590/S1517-838246220131206info:eu-repo/semantics/altIdentifier/url/http://ref.scielo.org/762b6sinfo:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4507559/info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:46:58Zoai:ri.conicet.gov.ar:11336/38587instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:46:58.495CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
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 D.
BRUCELLOSIS
LOOP-MEDIATED ISOTHERMAL AMPLIFICATION
MOLECULAR TYPING
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 D.
Gioffré, Andrea Karina
Cerón Cucchi, María Esperanza
Caimi, Karina Cynthia
Ruybal, Paula
Zumárraga, Martín José
Cravero, Silvio Lorenzo Pedro
author Trangoni, Marcos D.
author_facet Trangoni, Marcos D.
Gioffré, Andrea Karina
Cerón Cucchi, María Esperanza
Caimi, Karina Cynthia
Ruybal, Paula
Zumárraga, Martín José
Cravero, Silvio Lorenzo Pedro
author_role author
author2 Gioffré, Andrea Karina
Cerón Cucchi, María Esperanza
Caimi, Karina Cynthia
Ruybal, Paula
Zumárraga, Martín José
Cravero, Silvio Lorenzo Pedro
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv BRUCELLOSIS
LOOP-MEDIATED ISOTHERMAL AMPLIFICATION
MOLECULAR TYPING
PARATUBERCULOSIS
topic BRUCELLOSIS
LOOP-MEDIATED ISOTHERMAL AMPLIFICATION
MOLECULAR TYPING
PARATUBERCULOSIS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
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 Green ™ 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.
Fil: Trangoni, Marcos D.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Gioffré, Andrea Karina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Cerón Cucchi, María Esperanza. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Patobiología; Argentina
Fil: Caimi, Karina Cynthia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Ruybal, Paula. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Zumárraga, Martín José. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Cravero, Silvio Lorenzo Pedro. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. 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 Green ™ 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
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 http://hdl.handle.net/11336/38587
Trangoni, Marcos D.; Gioffré, Andrea Karina; Cerón Cucchi, María Esperanza; Caimi, Karina Cynthia; Ruybal, Paula; et al.; LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis; Sociedade Brasileira de Microbiologia; Brazilian Journal of Microbiology; 46; 2; 6-2015; 619-626
1517-8382
1678-4405
CONICET Digital
CONICET
url http://hdl.handle.net/11336/38587
identifier_str_mv Trangoni, Marcos D.; Gioffré, Andrea Karina; Cerón Cucchi, María Esperanza; Caimi, Karina Cynthia; Ruybal, Paula; et al.; LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis; Sociedade Brasileira de Microbiologia; Brazilian Journal of Microbiology; 46; 2; 6-2015; 619-626
1517-8382
1678-4405
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1590/S1517-838246220131206
info:eu-repo/semantics/altIdentifier/url/http://ref.scielo.org/762b6s
info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4507559/
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc/2.5/ar/
dc.format.none.fl_str_mv application/pdf
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 reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.069144

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