Detection of pathogenic leptospiral DNA traces in canine sera serum samples by loop mediated isothermal amplification (LAMP)

Autores
Grune Loffler, Sylvia; Leiva, Carlos; Scialfa, Exequiel; Redondo, Leandro Martin; Florin-Christensen, Mónica; Martinez, Mara Leila; Romero, Graciela Noemi; Brihuega, Bibiana Felicitas
Año de publicación
2016
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Leptospirosis is a worldwide distributed zoonosis, caused by pathogenic strains of spirochetes belonging to the genus Leptospira spp. So far, the only gold standard diagnostic method for this disease has been the Micro-Agglutination Test (MAT). The purpose of this study was to test whether a correlation exists between serological diagnosis by MAT and the presence of Leptospira spp. DNA traces in serum. Anti-Leptospira spp. antibody titers were determined in dog serum samples obtained at a canine shelter from Buenos Aires Province, Argentina (n= 80) by MAT using ten Leptospira spp. serovars. DNA extracted from these samples using Chelex-100 resin was used as template for Leptospira spp. specific PCR and loop-mediated isothermal amplification (LAMP) tests. In the latter case, product detection by incubation with malachite green dye was set up, obtaining optimal results at a dye concentration of 0.02% (w(v)) and a pH of 6.1. PCR and LAMP showed the same sensitivity, detecting leptospiral DNA in 12 out of the 48 MAT- positive sera. No detection was attained by either method in MAT-negative samples. These results show that it is possible to use LAMP for the detection of leptospirosis in non-human mammals. It also shows that, in the case of serum samples, MAT has higher diagnostic sensitivity than the currently available PCR and LAMP tests.
Instituto de Patobiología
Fil: Grune Loffler, Sylvia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina
Fil: Grune Loffler, Sylvia. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina
Fil: Leiva, Carlos. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina
Fil: Leiva, Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina
Fil: Scialfa, Exequiel. Ministerio de Salud de la Provincia de Buenos Aires. División Zoonosis Rurales; Argentina
Fil: Redondo, Leandro Martin. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina
Fil: Redondo, Leandro Martin. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina
Fil: Florin-Christensen, Mónica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina
Fil: Florin-Christensen, Mónica. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina
Fil: Martinez, Mara Leila. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina
Fil: Romero, Graciela Noemi. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina
Fil: Brihuega, Bibiana Felicitas. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina
Fuente
Immunology and Infectious Diseases 4 (4) : 39-43. (2016)
Materia
Leptospirosis
Técnica Inmunoenzimática
Proteína Sanguínea
Perros
Buenos Aires (provincia)
Immunoenzyme Techniques
Blood Proteins
Dogs
Buenos Aires (province)
Loop Mediated Isothermal Amplification (LAMP)
DNA Traces
Canine Sera
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:20.500.12123/26941
Acceder
id INTADig_b4a1d628a751e86ec836865687cfe93e
oai_identifier_str oai:localhost:20.500.12123/26941
network_acronym_str INTADig
repository_id_str l
network_name_str INTA Digital (INTA)
spelling Detection of pathogenic leptospiral DNA traces in canine sera serum samples by loop mediated isothermal amplification (LAMP)Grune Loffler, SylviaLeiva, CarlosScialfa, ExequielRedondo, Leandro MartinFlorin-Christensen, MónicaMartinez, Mara LeilaRomero, Graciela NoemiBrihuega, Bibiana FelicitasLeptospirosisTécnica InmunoenzimáticaProteína SanguíneaPerrosBuenos Aires (provincia)Immunoenzyme TechniquesBlood ProteinsDogsBuenos Aires (province)Loop Mediated Isothermal Amplification (LAMP)DNA TracesCanine SeraLeptospirosis is a worldwide distributed zoonosis, caused by pathogenic strains of spirochetes belonging to the genus Leptospira spp. So far, the only gold standard diagnostic method for this disease has been the Micro-Agglutination Test (MAT). The purpose of this study was to test whether a correlation exists between serological diagnosis by MAT and the presence of Leptospira spp. DNA traces in serum. Anti-Leptospira spp. antibody titers were determined in dog serum samples obtained at a canine shelter from Buenos Aires Province, Argentina (n= 80) by MAT using ten Leptospira spp. serovars. DNA extracted from these samples using Chelex-100 resin was used as template for Leptospira spp. specific PCR and loop-mediated isothermal amplification (LAMP) tests. In the latter case, product detection by incubation with malachite green dye was set up, obtaining optimal results at a dye concentration of 0.02% (w(v)) and a pH of 6.1. PCR and LAMP showed the same sensitivity, detecting leptospiral DNA in 12 out of the 48 MAT- positive sera. No detection was attained by either method in MAT-negative samples. These results show that it is possible to use LAMP for the detection of leptospirosis in non-human mammals. It also shows that, in the case of serum samples, MAT has higher diagnostic sensitivity than the currently available PCR and LAMP tests.Instituto de PatobiologíaFil: Grune Loffler, Sylvia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Grune Loffler, Sylvia. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); ArgentinaFil: Leiva, Carlos. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Leiva, Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); ArgentinaFil: Scialfa, Exequiel. Ministerio de Salud de la Provincia de Buenos Aires. División Zoonosis Rurales; ArgentinaFil: Redondo, Leandro Martin. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Redondo, Leandro Martin. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); ArgentinaFil: Florin-Christensen, Mónica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Florin-Christensen, Mónica. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); ArgentinaFil: Martinez, Mara Leila. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Romero, Graciela Noemi. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaFil: Brihuega, Bibiana Felicitas. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; ArgentinaHorizon Research2026-07-06T19:31:47Z2026-07-06T19:31:47Z2016info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/26941https://www.hrpub.org/journals/article_info.php?aid=54482333-2719 (impreso)2333-3286 (online)doi: https://doi.org/10.13189/iid.2016.040401Immunology and Infectious Diseases 4 (4) : 39-43. (2016)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo: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)2026-11-05T11:51:29Zoai:localhost:20.500.12123/26941instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2026-11-05 11:51:30.087INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Detection of pathogenic leptospiral DNA traces in canine sera serum samples by loop mediated isothermal amplification (LAMP)
title Detection of pathogenic leptospiral DNA traces in canine sera serum samples by loop mediated isothermal amplification (LAMP)
spellingShingle Detection of pathogenic leptospiral DNA traces in canine sera serum samples by loop mediated isothermal amplification (LAMP)
Grune Loffler, Sylvia
Leptospirosis
Técnica Inmunoenzimática
Proteína Sanguínea
Perros
Buenos Aires (provincia)
Immunoenzyme Techniques
Blood Proteins
Dogs
Buenos Aires (province)
Loop Mediated Isothermal Amplification (LAMP)
DNA Traces
Canine Sera
title_short Detection of pathogenic leptospiral DNA traces in canine sera serum samples by loop mediated isothermal amplification (LAMP)
title_full Detection of pathogenic leptospiral DNA traces in canine sera serum samples by loop mediated isothermal amplification (LAMP)
title_fullStr Detection of pathogenic leptospiral DNA traces in canine sera serum samples by loop mediated isothermal amplification (LAMP)
title_full_unstemmed Detection of pathogenic leptospiral DNA traces in canine sera serum samples by loop mediated isothermal amplification (LAMP)
title_sort Detection of pathogenic leptospiral DNA traces in canine sera serum samples by loop mediated isothermal amplification (LAMP)
dc.creator.none.fl_str_mv Grune Loffler, Sylvia
Leiva, Carlos
Scialfa, Exequiel
Redondo, Leandro Martin
Florin-Christensen, Mónica
Martinez, Mara Leila
Romero, Graciela Noemi
Brihuega, Bibiana Felicitas
author Grune Loffler, Sylvia
author_facet Grune Loffler, Sylvia
Leiva, Carlos
Scialfa, Exequiel
Redondo, Leandro Martin
Florin-Christensen, Mónica
Martinez, Mara Leila
Romero, Graciela Noemi
Brihuega, Bibiana Felicitas
author_role author
author2 Leiva, Carlos
Scialfa, Exequiel
Redondo, Leandro Martin
Florin-Christensen, Mónica
Martinez, Mara Leila
Romero, Graciela Noemi
Brihuega, Bibiana Felicitas
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Leptospirosis
Técnica Inmunoenzimática
Proteína Sanguínea
Perros
Buenos Aires (provincia)
Immunoenzyme Techniques
Blood Proteins
Dogs
Buenos Aires (province)
Loop Mediated Isothermal Amplification (LAMP)
DNA Traces
Canine Sera
topic Leptospirosis
Técnica Inmunoenzimática
Proteína Sanguínea
Perros
Buenos Aires (provincia)
Immunoenzyme Techniques
Blood Proteins
Dogs
Buenos Aires (province)
Loop Mediated Isothermal Amplification (LAMP)
DNA Traces
Canine Sera
dc.description.none.fl_txt_mv Leptospirosis is a worldwide distributed zoonosis, caused by pathogenic strains of spirochetes belonging to the genus Leptospira spp. So far, the only gold standard diagnostic method for this disease has been the Micro-Agglutination Test (MAT). The purpose of this study was to test whether a correlation exists between serological diagnosis by MAT and the presence of Leptospira spp. DNA traces in serum. Anti-Leptospira spp. antibody titers were determined in dog serum samples obtained at a canine shelter from Buenos Aires Province, Argentina (n= 80) by MAT using ten Leptospira spp. serovars. DNA extracted from these samples using Chelex-100 resin was used as template for Leptospira spp. specific PCR and loop-mediated isothermal amplification (LAMP) tests. In the latter case, product detection by incubation with malachite green dye was set up, obtaining optimal results at a dye concentration of 0.02% (w(v)) and a pH of 6.1. PCR and LAMP showed the same sensitivity, detecting leptospiral DNA in 12 out of the 48 MAT- positive sera. No detection was attained by either method in MAT-negative samples. These results show that it is possible to use LAMP for the detection of leptospirosis in non-human mammals. It also shows that, in the case of serum samples, MAT has higher diagnostic sensitivity than the currently available PCR and LAMP tests.
Instituto de Patobiología
Fil: Grune Loffler, Sylvia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina
Fil: Grune Loffler, Sylvia. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina
Fil: Leiva, Carlos. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina
Fil: Leiva, Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina
Fil: Scialfa, Exequiel. Ministerio de Salud de la Provincia de Buenos Aires. División Zoonosis Rurales; Argentina
Fil: Redondo, Leandro Martin. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina
Fil: Redondo, Leandro Martin. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina
Fil: Florin-Christensen, Mónica. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina
Fil: Florin-Christensen, Mónica. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina
Fil: Martinez, Mara Leila. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina
Fil: Romero, Graciela Noemi. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina
Fil: Brihuega, Bibiana Felicitas. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología; Argentina
description Leptospirosis is a worldwide distributed zoonosis, caused by pathogenic strains of spirochetes belonging to the genus Leptospira spp. So far, the only gold standard diagnostic method for this disease has been the Micro-Agglutination Test (MAT). The purpose of this study was to test whether a correlation exists between serological diagnosis by MAT and the presence of Leptospira spp. DNA traces in serum. Anti-Leptospira spp. antibody titers were determined in dog serum samples obtained at a canine shelter from Buenos Aires Province, Argentina (n= 80) by MAT using ten Leptospira spp. serovars. DNA extracted from these samples using Chelex-100 resin was used as template for Leptospira spp. specific PCR and loop-mediated isothermal amplification (LAMP) tests. In the latter case, product detection by incubation with malachite green dye was set up, obtaining optimal results at a dye concentration of 0.02% (w(v)) and a pH of 6.1. PCR and LAMP showed the same sensitivity, detecting leptospiral DNA in 12 out of the 48 MAT- positive sera. No detection was attained by either method in MAT-negative samples. These results show that it is possible to use LAMP for the detection of leptospirosis in non-human mammals. It also shows that, in the case of serum samples, MAT has higher diagnostic sensitivity than the currently available PCR and LAMP tests.
publishDate 2016
dc.date.none.fl_str_mv 2016
2026-07-06T19:31:47Z
2026-07-06T19:31:47Z
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/20.500.12123/26941
https://www.hrpub.org/journals/article_info.php?aid=5448
2333-2719 (impreso)
2333-3286 (online)
doi: https://doi.org/10.13189/iid.2016.040401
url http://hdl.handle.net/20.500.12123/26941
https://www.hrpub.org/journals/article_info.php?aid=5448
identifier_str_mv 2333-2719 (impreso)
2333-3286 (online)
doi: https://doi.org/10.13189/iid.2016.040401
dc.language.none.fl_str_mv eng
language eng
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 Horizon Research
publisher.none.fl_str_mv Horizon Research
dc.source.none.fl_str_mv Immunology and Infectious Diseases 4 (4) : 39-43. (2016)
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
_version_ 1881032524263587840
score 12.823554

Publicaciones similares