Enhanced detection of Leptospira in cattle : Comparative performance of loop-mediated isothermal amplification, polymerase chain reaction and serological methods

Autores
Hamer, Micaela; Saraullo, Vanina Rosa; Esteban, Micaela; Sanchez, Maria Cristina; Brihuega, Bibiana Felicitas; Martinez, Mara Leila
Año de publicación
2025
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Leptospirosis is a globally distributed zoonotic disease. This study aimed to evaluate the performance of loop-mediated isothermal amplification (LAMP) in detecting Leptospira DNA in bovine samples compared to conventional lipL32 PCR and serological methods. A total of 464 serum, 96 urine, and 31 organ samples were analyzed using LAMP, lipL32 PCR, and the microscopic agglutination test (MAT). The results showed that 52.8 % of sera tested positive exclusively by MAT, indicating past exposure, while 1.2 % tested positive by molecular techniques, suggesting early infection. In urine samples, LAMP detected leptospiral DNA in 62.5 % of cases, highlighting its potential for identifying asymptomatic carriers. Among organ samples from aborted fetuses, 22.6 % were positive by both molecular techniques, supporting Leptospira as a potential cause of fetal loss. Notably, LAMP exhibited a high diagnostic sensitivity (100 %, 95 % CI: 93.5–100 %) and specificity (95.4 %, 95 % CI: 93.2–97.0 %), outperforming lipL32 PCR in DNA detection. Additionally, Cohen’s Kappa index (0.792) indicated substantial agreement between LAMP and lipL32 PCR. Given its higher sensitivity, rapid turnaround, and affordability, LAMP represents a valuable tool for improving leptospirosis diagnosis, particularly in resource-limited settings. These findings underscore the importance of integrating molecular and serological techniques for a more accurate detection of Leptospira infection in cattle.
Instituto de Patobiología
Fil: Hamer, Micaela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina
Fil: Hamer, Micaela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Saraullo, Vanina Rosa. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina
Fil: Saraullo, Vanina Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Esteban, Micaela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina
Fil: Esteban, Micaela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Sanchez, Maria Cristina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina
Fil: Sanchez, Maria Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Brihuega, Bibiana Felicitas. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina
Fil: Brihuega, Bibiana Felicitas. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Brihuega, Bibiana Felicitas. Universidad del Salvador. Escuela de Veterinaria; Argentina
Fil: Martinez, Mara Leila. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina
Fil: Martinez, Mara Leila. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Martinez, Mara Leila. Universidad del Salvador. Escuela de Veterinaria; Argentina
Fuente
Veterinary Microbiology 308 : 110662 (September 2025)
Materia
PCR
Agglutination Tests
Leptospirosis
Cattle
Isothermal Processes
Immunological Techniques
Leptospira
Reacción de Aglutinación
Ganado Bovino
Proceso Isotérmico
Técnicas Inmunológicas
Nivel de accesibilidad
acceso restringido
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:20.500.12123/24460
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oai_identifier_str oai:localhost:20.500.12123/24460
network_acronym_str INTADig
repository_id_str l
network_name_str INTA Digital (INTA)
spelling Enhanced detection of Leptospira in cattle : Comparative performance of loop-mediated isothermal amplification, polymerase chain reaction and serological methodsHamer, MicaelaSaraullo, Vanina RosaEsteban, MicaelaSanchez, Maria CristinaBrihuega, Bibiana FelicitasMartinez, Mara LeilaPCRAgglutination TestsLeptospirosisCattleIsothermal ProcessesImmunological TechniquesLeptospiraReacción de AglutinaciónGanado BovinoProceso IsotérmicoTécnicas InmunológicasLeptospirosis is a globally distributed zoonotic disease. This study aimed to evaluate the performance of loop-mediated isothermal amplification (LAMP) in detecting Leptospira DNA in bovine samples compared to conventional lipL32 PCR and serological methods. A total of 464 serum, 96 urine, and 31 organ samples were analyzed using LAMP, lipL32 PCR, and the microscopic agglutination test (MAT). The results showed that 52.8 % of sera tested positive exclusively by MAT, indicating past exposure, while 1.2 % tested positive by molecular techniques, suggesting early infection. In urine samples, LAMP detected leptospiral DNA in 62.5 % of cases, highlighting its potential for identifying asymptomatic carriers. Among organ samples from aborted fetuses, 22.6 % were positive by both molecular techniques, supporting Leptospira as a potential cause of fetal loss. Notably, LAMP exhibited a high diagnostic sensitivity (100 %, 95 % CI: 93.5–100 %) and specificity (95.4 %, 95 % CI: 93.2–97.0 %), outperforming lipL32 PCR in DNA detection. Additionally, Cohen’s Kappa index (0.792) indicated substantial agreement between LAMP and lipL32 PCR. Given its higher sensitivity, rapid turnaround, and affordability, LAMP represents a valuable tool for improving leptospirosis diagnosis, particularly in resource-limited settings. These findings underscore the importance of integrating molecular and serological techniques for a more accurate detection of Leptospira infection in cattle.Instituto de PatobiologíaFil: Hamer, Micaela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; ArgentinaFil: Hamer, Micaela. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Saraullo, Vanina Rosa. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; ArgentinaFil: Saraullo, Vanina Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Esteban, Micaela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; ArgentinaFil: Esteban, Micaela. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Sanchez, Maria Cristina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; ArgentinaFil: Sanchez, Maria Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Brihuega, Bibiana Felicitas. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; ArgentinaFil: Brihuega, Bibiana Felicitas. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Brihuega, Bibiana Felicitas. Universidad del Salvador. Escuela de Veterinaria; ArgentinaFil: Martinez, Mara Leila. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; ArgentinaFil: Martinez, Mara Leila. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Martinez, Mara Leila. Universidad del Salvador. Escuela de Veterinaria; ArgentinaElsevier2025-11-05T11:16:47Z2025-11-05T11:16:47Z2025-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/24460https://www.sciencedirect.com/science/article/abs/pii/S03781135250029740378-1135https://doi.org/10.1016/j.vetmic.2025.110662Veterinary Microbiology 308 : 110662 (September 2025)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/restrictedAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2025-11-27T08:40:52Zoai:localhost:20.500.12123/24460instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-11-27 08:40:52.853INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Enhanced detection of Leptospira in cattle : Comparative performance of loop-mediated isothermal amplification, polymerase chain reaction and serological methods
title Enhanced detection of Leptospira in cattle : Comparative performance of loop-mediated isothermal amplification, polymerase chain reaction and serological methods
spellingShingle Enhanced detection of Leptospira in cattle : Comparative performance of loop-mediated isothermal amplification, polymerase chain reaction and serological methods
Hamer, Micaela
PCR
Agglutination Tests
Leptospirosis
Cattle
Isothermal Processes
Immunological Techniques
Leptospira
Reacción de Aglutinación
Ganado Bovino
Proceso Isotérmico
Técnicas Inmunológicas
title_short Enhanced detection of Leptospira in cattle : Comparative performance of loop-mediated isothermal amplification, polymerase chain reaction and serological methods
title_full Enhanced detection of Leptospira in cattle : Comparative performance of loop-mediated isothermal amplification, polymerase chain reaction and serological methods
title_fullStr Enhanced detection of Leptospira in cattle : Comparative performance of loop-mediated isothermal amplification, polymerase chain reaction and serological methods
title_full_unstemmed Enhanced detection of Leptospira in cattle : Comparative performance of loop-mediated isothermal amplification, polymerase chain reaction and serological methods
title_sort Enhanced detection of Leptospira in cattle : Comparative performance of loop-mediated isothermal amplification, polymerase chain reaction and serological methods
dc.creator.none.fl_str_mv Hamer, Micaela
Saraullo, Vanina Rosa
Esteban, Micaela
Sanchez, Maria Cristina
Brihuega, Bibiana Felicitas
Martinez, Mara Leila
author Hamer, Micaela
author_facet Hamer, Micaela
Saraullo, Vanina Rosa
Esteban, Micaela
Sanchez, Maria Cristina
Brihuega, Bibiana Felicitas
Martinez, Mara Leila
author_role author
author2 Saraullo, Vanina Rosa
Esteban, Micaela
Sanchez, Maria Cristina
Brihuega, Bibiana Felicitas
Martinez, Mara Leila
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv PCR
Agglutination Tests
Leptospirosis
Cattle
Isothermal Processes
Immunological Techniques
Leptospira
Reacción de Aglutinación
Ganado Bovino
Proceso Isotérmico
Técnicas Inmunológicas
topic PCR
Agglutination Tests
Leptospirosis
Cattle
Isothermal Processes
Immunological Techniques
Leptospira
Reacción de Aglutinación
Ganado Bovino
Proceso Isotérmico
Técnicas Inmunológicas
dc.description.none.fl_txt_mv Leptospirosis is a globally distributed zoonotic disease. This study aimed to evaluate the performance of loop-mediated isothermal amplification (LAMP) in detecting Leptospira DNA in bovine samples compared to conventional lipL32 PCR and serological methods. A total of 464 serum, 96 urine, and 31 organ samples were analyzed using LAMP, lipL32 PCR, and the microscopic agglutination test (MAT). The results showed that 52.8 % of sera tested positive exclusively by MAT, indicating past exposure, while 1.2 % tested positive by molecular techniques, suggesting early infection. In urine samples, LAMP detected leptospiral DNA in 62.5 % of cases, highlighting its potential for identifying asymptomatic carriers. Among organ samples from aborted fetuses, 22.6 % were positive by both molecular techniques, supporting Leptospira as a potential cause of fetal loss. Notably, LAMP exhibited a high diagnostic sensitivity (100 %, 95 % CI: 93.5–100 %) and specificity (95.4 %, 95 % CI: 93.2–97.0 %), outperforming lipL32 PCR in DNA detection. Additionally, Cohen’s Kappa index (0.792) indicated substantial agreement between LAMP and lipL32 PCR. Given its higher sensitivity, rapid turnaround, and affordability, LAMP represents a valuable tool for improving leptospirosis diagnosis, particularly in resource-limited settings. These findings underscore the importance of integrating molecular and serological techniques for a more accurate detection of Leptospira infection in cattle.
Instituto de Patobiología
Fil: Hamer, Micaela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina
Fil: Hamer, Micaela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Saraullo, Vanina Rosa. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina
Fil: Saraullo, Vanina Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Esteban, Micaela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina
Fil: Esteban, Micaela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Sanchez, Maria Cristina. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina
Fil: Sanchez, Maria Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Brihuega, Bibiana Felicitas. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina
Fil: Brihuega, Bibiana Felicitas. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Brihuega, Bibiana Felicitas. Universidad del Salvador. Escuela de Veterinaria; Argentina
Fil: Martinez, Mara Leila. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina
Fil: Martinez, Mara Leila. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Martinez, Mara Leila. Universidad del Salvador. Escuela de Veterinaria; Argentina
description Leptospirosis is a globally distributed zoonotic disease. This study aimed to evaluate the performance of loop-mediated isothermal amplification (LAMP) in detecting Leptospira DNA in bovine samples compared to conventional lipL32 PCR and serological methods. A total of 464 serum, 96 urine, and 31 organ samples were analyzed using LAMP, lipL32 PCR, and the microscopic agglutination test (MAT). The results showed that 52.8 % of sera tested positive exclusively by MAT, indicating past exposure, while 1.2 % tested positive by molecular techniques, suggesting early infection. In urine samples, LAMP detected leptospiral DNA in 62.5 % of cases, highlighting its potential for identifying asymptomatic carriers. Among organ samples from aborted fetuses, 22.6 % were positive by both molecular techniques, supporting Leptospira as a potential cause of fetal loss. Notably, LAMP exhibited a high diagnostic sensitivity (100 %, 95 % CI: 93.5–100 %) and specificity (95.4 %, 95 % CI: 93.2–97.0 %), outperforming lipL32 PCR in DNA detection. Additionally, Cohen’s Kappa index (0.792) indicated substantial agreement between LAMP and lipL32 PCR. Given its higher sensitivity, rapid turnaround, and affordability, LAMP represents a valuable tool for improving leptospirosis diagnosis, particularly in resource-limited settings. These findings underscore the importance of integrating molecular and serological techniques for a more accurate detection of Leptospira infection in cattle.
publishDate 2025
dc.date.none.fl_str_mv 2025-11-05T11:16:47Z
2025-11-05T11:16:47Z
2025-09
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/20.500.12123/24460
https://www.sciencedirect.com/science/article/abs/pii/S0378113525002974
0378-1135
https://doi.org/10.1016/j.vetmic.2025.110662
url http://hdl.handle.net/20.500.12123/24460
https://www.sciencedirect.com/science/article/abs/pii/S0378113525002974
https://doi.org/10.1016/j.vetmic.2025.110662
identifier_str_mv 0378-1135
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/restrictedAccess
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 restrictedAccess
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 Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv Veterinary Microbiology 308 : 110662 (September 2025)
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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