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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/38587
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oai:ri.conicet.gov.ar:11336/38587 |
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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 Técnicas2025-09-03T10:11:50Zoai: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:34982025-09-03 10:11:50.521CONICET 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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1842270172408184832 |
score |
13.13397 |