Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters

Autores
Hamer, Micaela; Watanabe, Olivia; Saraullo, Vanina Rosa; Ortega, Facundo Miguel; Sanchez, Maria Cristina; Martinez, Mara Leila; Brihuega, Bibiana Felicitas; Grune Loffler, Sylvia
Año de publicación
2023
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Leptospirosis is a zoonotic disease with significant public health and economic impact worldwide. Rapid and accurate diagnosis is essential for effective prevention and treatment. This study optimized a loop-mediated isothermal amplification (LAMP) assay using BFo isothermal DNA polymerase with different colorimetric indicators. LAMP was able to detect DNA from pathogenic and intermediate leptospires, while non-pathogenic leptospires and other non-leptospiral microorganisms were negative. LAMP assay combined with calcein showed a tenfold higher limit of detection (1 ng of leptospiral DNA per reaction) than LAMP combined with hydroxynaphthol blue or end-point PCR lipL32 (10 ng of DNA per reaction). Animal samples were collected from infected and non-infected Golden Syrian hamsters (Mesocricetus auratus) to evaluate and compare the performance of LAMP and PCR. These techniques showed a substantial agreement according to Cohen’s kappa statistic, being both useful techniques for detecting leptospiral DNA in clinical samples. Overall, this study demonstrates that the LAMP assay is a sensitive, specific, rapid, and simple tool for the detection of leptospiral DNA. It has the potential to facilitate the diagnosis of leptospirosis, particularly in low-income regions with limited diagnosis resources.
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: Watanabe, Olivia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina
Fil: Watanabe, Olivia. 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: Ortega, Facundo Miguel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina
Fil: Ortega, Facundo Miguel. 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: 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: 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: Grune Loffler, Sylvia. Universidad Abierta Interamericana (UAI); Argentina
Fil: Grune Loffler, Sylvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Virología Humana y Animal (CEVHAN); Argentina
Fuente
Veterinary Research Communications : 103-111 (2024)
Materia
Animal Models
Infectious Diseases
Leptospira
PCR
Hamsters
Modelos Animales
Enfermedades Infecciosas
Reacción en Cadena de la Polimerasa
Hámster
Loop-mediated Isothermal Amplification
Sensitivity
Specificity
Amplificación Isotérmica Mediada por Bucle
Sensibilidad
Especificidad
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:20.500.12123/15227
Acceder
id INTADig_60b4bbf1b322253c865482b0064e4f3a
oai_identifier_str oai:localhost:20.500.12123/15227
network_acronym_str INTADig
repository_id_str l
network_name_str INTA Digital (INTA)
spelling Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamstersHamer, MicaelaWatanabe, OliviaSaraullo, Vanina RosaOrtega, Facundo MiguelSanchez, Maria CristinaMartinez, Mara LeilaBrihuega, Bibiana FelicitasGrune Loffler, SylviaAnimal ModelsInfectious DiseasesLeptospiraPCRHamstersModelos AnimalesEnfermedades InfecciosasReacción en Cadena de la PolimerasaHámsterLoop-mediated Isothermal AmplificationSensitivitySpecificityAmplificación Isotérmica Mediada por BucleSensibilidadEspecificidadLeptospirosis is a zoonotic disease with significant public health and economic impact worldwide. Rapid and accurate diagnosis is essential for effective prevention and treatment. This study optimized a loop-mediated isothermal amplification (LAMP) assay using BFo isothermal DNA polymerase with different colorimetric indicators. LAMP was able to detect DNA from pathogenic and intermediate leptospires, while non-pathogenic leptospires and other non-leptospiral microorganisms were negative. LAMP assay combined with calcein showed a tenfold higher limit of detection (1 ng of leptospiral DNA per reaction) than LAMP combined with hydroxynaphthol blue or end-point PCR lipL32 (10 ng of DNA per reaction). Animal samples were collected from infected and non-infected Golden Syrian hamsters (Mesocricetus auratus) to evaluate and compare the performance of LAMP and PCR. These techniques showed a substantial agreement according to Cohen’s kappa statistic, being both useful techniques for detecting leptospiral DNA in clinical samples. Overall, this study demonstrates that the LAMP assay is a sensitive, specific, rapid, and simple tool for the detection of leptospiral DNA. It has the potential to facilitate the diagnosis of leptospirosis, particularly in low-income regions with limited diagnosis resources.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: Watanabe, Olivia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; ArgentinaFil: Watanabe, Olivia. 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: Ortega, Facundo Miguel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; ArgentinaFil: Ortega, Facundo Miguel. 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: 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: 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: Grune Loffler, Sylvia. Universidad Abierta Interamericana (UAI); ArgentinaFil: Grune Loffler, Sylvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Virología Humana y Animal (CEVHAN); ArgentinaSpringer2023-09-15T12:38:41Z2023-09-15T12:38:41Z2024-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/15227https://link.springer.com/article/10.1007/s11259-023-10183-11573-7446https://doi.org/10.1007/s11259-023-10183-1Veterinary Research Communications : 103-111 (2024)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)2025-09-29T13:46:06Zoai:localhost:20.500.12123/15227instacron: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 13:46:06.938INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters
title Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters
spellingShingle Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters
Hamer, Micaela
Animal Models
Infectious Diseases
Leptospira
PCR
Hamsters
Modelos Animales
Enfermedades Infecciosas
Reacción en Cadena de la Polimerasa
Hámster
Loop-mediated Isothermal Amplification
Sensitivity
Specificity
Amplificación Isotérmica Mediada por Bucle
Sensibilidad
Especificidad
title_short Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters
title_full Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters
title_fullStr Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters
title_full_unstemmed Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters
title_sort Optimization and comparative analysis of LAMP and PCR techniques for the detection of leptospiral DNA in Golden Syrian hamsters
dc.creator.none.fl_str_mv Hamer, Micaela
Watanabe, Olivia
Saraullo, Vanina Rosa
Ortega, Facundo Miguel
Sanchez, Maria Cristina
Martinez, Mara Leila
Brihuega, Bibiana Felicitas
Grune Loffler, Sylvia
author Hamer, Micaela
author_facet Hamer, Micaela
Watanabe, Olivia
Saraullo, Vanina Rosa
Ortega, Facundo Miguel
Sanchez, Maria Cristina
Martinez, Mara Leila
Brihuega, Bibiana Felicitas
Grune Loffler, Sylvia
author_role author
author2 Watanabe, Olivia
Saraullo, Vanina Rosa
Ortega, Facundo Miguel
Sanchez, Maria Cristina
Martinez, Mara Leila
Brihuega, Bibiana Felicitas
Grune Loffler, Sylvia
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Animal Models
Infectious Diseases
Leptospira
PCR
Hamsters
Modelos Animales
Enfermedades Infecciosas
Reacción en Cadena de la Polimerasa
Hámster
Loop-mediated Isothermal Amplification
Sensitivity
Specificity
Amplificación Isotérmica Mediada por Bucle
Sensibilidad
Especificidad
topic Animal Models
Infectious Diseases
Leptospira
PCR
Hamsters
Modelos Animales
Enfermedades Infecciosas
Reacción en Cadena de la Polimerasa
Hámster
Loop-mediated Isothermal Amplification
Sensitivity
Specificity
Amplificación Isotérmica Mediada por Bucle
Sensibilidad
Especificidad
dc.description.none.fl_txt_mv Leptospirosis is a zoonotic disease with significant public health and economic impact worldwide. Rapid and accurate diagnosis is essential for effective prevention and treatment. This study optimized a loop-mediated isothermal amplification (LAMP) assay using BFo isothermal DNA polymerase with different colorimetric indicators. LAMP was able to detect DNA from pathogenic and intermediate leptospires, while non-pathogenic leptospires and other non-leptospiral microorganisms were negative. LAMP assay combined with calcein showed a tenfold higher limit of detection (1 ng of leptospiral DNA per reaction) than LAMP combined with hydroxynaphthol blue or end-point PCR lipL32 (10 ng of DNA per reaction). Animal samples were collected from infected and non-infected Golden Syrian hamsters (Mesocricetus auratus) to evaluate and compare the performance of LAMP and PCR. These techniques showed a substantial agreement according to Cohen’s kappa statistic, being both useful techniques for detecting leptospiral DNA in clinical samples. Overall, this study demonstrates that the LAMP assay is a sensitive, specific, rapid, and simple tool for the detection of leptospiral DNA. It has the potential to facilitate the diagnosis of leptospirosis, particularly in low-income regions with limited diagnosis resources.
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: Watanabe, Olivia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina
Fil: Watanabe, Olivia. 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: Ortega, Facundo Miguel. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria. Laboratorio de Leptospirosis; Argentina
Fil: Ortega, Facundo Miguel. 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: 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: 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: Grune Loffler, Sylvia. Universidad Abierta Interamericana (UAI); Argentina
Fil: Grune Loffler, Sylvia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Virología Humana y Animal (CEVHAN); Argentina
description Leptospirosis is a zoonotic disease with significant public health and economic impact worldwide. Rapid and accurate diagnosis is essential for effective prevention and treatment. This study optimized a loop-mediated isothermal amplification (LAMP) assay using BFo isothermal DNA polymerase with different colorimetric indicators. LAMP was able to detect DNA from pathogenic and intermediate leptospires, while non-pathogenic leptospires and other non-leptospiral microorganisms were negative. LAMP assay combined with calcein showed a tenfold higher limit of detection (1 ng of leptospiral DNA per reaction) than LAMP combined with hydroxynaphthol blue or end-point PCR lipL32 (10 ng of DNA per reaction). Animal samples were collected from infected and non-infected Golden Syrian hamsters (Mesocricetus auratus) to evaluate and compare the performance of LAMP and PCR. These techniques showed a substantial agreement according to Cohen’s kappa statistic, being both useful techniques for detecting leptospiral DNA in clinical samples. Overall, this study demonstrates that the LAMP assay is a sensitive, specific, rapid, and simple tool for the detection of leptospiral DNA. It has the potential to facilitate the diagnosis of leptospirosis, particularly in low-income regions with limited diagnosis resources.
publishDate 2023
dc.date.none.fl_str_mv 2023-09-15T12:38:41Z
2023-09-15T12:38:41Z
2024-03
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/20.500.12123/15227
https://link.springer.com/article/10.1007/s11259-023-10183-1
1573-7446
https://doi.org/10.1007/s11259-023-10183-1
url http://hdl.handle.net/20.500.12123/15227
https://link.springer.com/article/10.1007/s11259-023-10183-1
https://doi.org/10.1007/s11259-023-10183-1
identifier_str_mv 1573-7446
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 Springer
publisher.none.fl_str_mv Springer
dc.source.none.fl_str_mv Veterinary Research Communications : 103-111 (2024)
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_ 1844619179161812992
score 12.559606

Publicaciones similares