Development and evaluation of a TaqMan® real-time PCR assay for species-specific detection of Ehrlichia canis

Autores
Sarli, Macarena; De Salvo, María N.; Díaz Pérez, Paula M.; Cicuttin, Gabriel L.; Nava, Santiago; Sebastian, Patrick
Año de publicación
2024
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The aim of this work was to develop a real-time PCR assay with a TaqMan® probe that detects a species-specific part of the 16S rDNA gene of Ehrlichia canis. Canine blood samples (n = 207), collected and tested by a conventional PCR assay within a study conducted by De Salvo et al., were simultaneously analyzed with the novel designed real-time PCR, and the results of both assays were compared. The agreement between the two methods was 97.6 % with a kappa value of 0.92186. Hereby, the standard error was 0.034416 and the 95 % confidence interval from 0.8544 to 0.98931. While the conventional PCR assay showed false negative results (2.42 %; 5/207), the real-time PCR assays showed a specificity of 100 %. The results of the current study showed that the developed assay presents sensitivity and specificity for the detection of E. canis in blood samples, adding a new tool for the diagnosis of this pathogen.
Asociacion Cooperadora INTA Rafaela. CONICET (Consejo Nacional de Investigaciones Científicas y Tecnicas).
EEA Rafaela
Fil: Sarli, Macarena. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Sarli, Macarena. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: De Salvo, M.N. Instituto de Zoonosis Luis Pasteur; Argentina
Fil: Díaz Pérez, Paula M. Instituto de Zoonosis Luis Pasteur; Argentina
Fil: Cicuttin, Gabriel L. Instituto de Zoonosis Luis Pasteur; Argentina
Fil: Nava, Santiago. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Nava, Santiago. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Sebastian, Patrick. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Sebastian, Patrick. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fuente
Diagnostic Microbiology and Infectious Disease 110 (4) : 116517 (December 2024)
Materia
Tick-borne Diseases
Diagnosis
Veterinary Medicine
Dogs
Enfermedad Transmitida por Garrapatas
Diagnóstico
Medicina Veterinaria
Ehrlichia canis
Perro
Hemoparasite
Hemoparásitos
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:20.500.12123/19510
Acceder
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oai_identifier_str oai:localhost:20.500.12123/19510
network_acronym_str INTADig
repository_id_str l
network_name_str INTA Digital (INTA)
spelling Development and evaluation of a TaqMan® real-time PCR assay for species-specific detection of Ehrlichia canisSarli, MacarenaDe Salvo, María N.Díaz Pérez, Paula M.Cicuttin, Gabriel L.Nava, SantiagoSebastian, PatrickTick-borne DiseasesDiagnosisVeterinary MedicineDogsEnfermedad Transmitida por GarrapatasDiagnósticoMedicina VeterinariaEhrlichia canisPerroHemoparasiteHemoparásitosThe aim of this work was to develop a real-time PCR assay with a TaqMan® probe that detects a species-specific part of the 16S rDNA gene of Ehrlichia canis. Canine blood samples (n = 207), collected and tested by a conventional PCR assay within a study conducted by De Salvo et al., were simultaneously analyzed with the novel designed real-time PCR, and the results of both assays were compared. The agreement between the two methods was 97.6 % with a kappa value of 0.92186. Hereby, the standard error was 0.034416 and the 95 % confidence interval from 0.8544 to 0.98931. While the conventional PCR assay showed false negative results (2.42 %; 5/207), the real-time PCR assays showed a specificity of 100 %. The results of the current study showed that the developed assay presents sensitivity and specificity for the detection of E. canis in blood samples, adding a new tool for the diagnosis of this pathogen.Asociacion Cooperadora INTA Rafaela. CONICET (Consejo Nacional de Investigaciones Científicas y Tecnicas).EEA RafaelaFil: Sarli, Macarena. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); ArgentinaFil: Sarli, Macarena. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); ArgentinaFil: De Salvo, M.N. Instituto de Zoonosis Luis Pasteur; ArgentinaFil: Díaz Pérez, Paula M. Instituto de Zoonosis Luis Pasteur; ArgentinaFil: Cicuttin, Gabriel L. Instituto de Zoonosis Luis Pasteur; ArgentinaFil: Nava, Santiago. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); ArgentinaFil: Nava, Santiago. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); ArgentinaFil: Sebastian, Patrick. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); ArgentinaFil: Sebastian, Patrick. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); ArgentinaElsevier2024-09-24T10:32:37Z2024-09-24T10:32:37Z2024-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/19510https://www.sciencedirect.com/science/article/pii/S07328893240034320732-88931879-0070https://doi.org/10.1016/j.diagmicrobio.2024.116517Diagnostic Microbiology and Infectious Disease 110 (4) : 116517 (December 2024)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repograntAgreement/INTA/2019-PE-E5-I109-001, Convocatoria: Estudios para el control de enfermedades subtropicales y/o transmitidas por vectores (Tristeza Bovina, Garrapatas, Miasis, Tripanosomiasis, Lengua Azul y lainfo: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)2025-09-04T09:50:37Zoai:localhost:20.500.12123/19510instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-04 09:50:37.738INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Development and evaluation of a TaqMan® real-time PCR assay for species-specific detection of Ehrlichia canis
title Development and evaluation of a TaqMan® real-time PCR assay for species-specific detection of Ehrlichia canis
spellingShingle Development and evaluation of a TaqMan® real-time PCR assay for species-specific detection of Ehrlichia canis
Sarli, Macarena
Tick-borne Diseases
Diagnosis
Veterinary Medicine
Dogs
Enfermedad Transmitida por Garrapatas
Diagnóstico
Medicina Veterinaria
Ehrlichia canis
Perro
Hemoparasite
Hemoparásitos
title_short Development and evaluation of a TaqMan® real-time PCR assay for species-specific detection of Ehrlichia canis
title_full Development and evaluation of a TaqMan® real-time PCR assay for species-specific detection of Ehrlichia canis
title_fullStr Development and evaluation of a TaqMan® real-time PCR assay for species-specific detection of Ehrlichia canis
title_full_unstemmed Development and evaluation of a TaqMan® real-time PCR assay for species-specific detection of Ehrlichia canis
title_sort Development and evaluation of a TaqMan® real-time PCR assay for species-specific detection of Ehrlichia canis
dc.creator.none.fl_str_mv Sarli, Macarena
De Salvo, María N.
Díaz Pérez, Paula M.
Cicuttin, Gabriel L.
Nava, Santiago
Sebastian, Patrick
author Sarli, Macarena
author_facet Sarli, Macarena
De Salvo, María N.
Díaz Pérez, Paula M.
Cicuttin, Gabriel L.
Nava, Santiago
Sebastian, Patrick
author_role author
author2 De Salvo, María N.
Díaz Pérez, Paula M.
Cicuttin, Gabriel L.
Nava, Santiago
Sebastian, Patrick
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Tick-borne Diseases
Diagnosis
Veterinary Medicine
Dogs
Enfermedad Transmitida por Garrapatas
Diagnóstico
Medicina Veterinaria
Ehrlichia canis
Perro
Hemoparasite
Hemoparásitos
topic Tick-borne Diseases
Diagnosis
Veterinary Medicine
Dogs
Enfermedad Transmitida por Garrapatas
Diagnóstico
Medicina Veterinaria
Ehrlichia canis
Perro
Hemoparasite
Hemoparásitos
dc.description.none.fl_txt_mv The aim of this work was to develop a real-time PCR assay with a TaqMan® probe that detects a species-specific part of the 16S rDNA gene of Ehrlichia canis. Canine blood samples (n = 207), collected and tested by a conventional PCR assay within a study conducted by De Salvo et al., were simultaneously analyzed with the novel designed real-time PCR, and the results of both assays were compared. The agreement between the two methods was 97.6 % with a kappa value of 0.92186. Hereby, the standard error was 0.034416 and the 95 % confidence interval from 0.8544 to 0.98931. While the conventional PCR assay showed false negative results (2.42 %; 5/207), the real-time PCR assays showed a specificity of 100 %. The results of the current study showed that the developed assay presents sensitivity and specificity for the detection of E. canis in blood samples, adding a new tool for the diagnosis of this pathogen.
Asociacion Cooperadora INTA Rafaela. CONICET (Consejo Nacional de Investigaciones Científicas y Tecnicas).
EEA Rafaela
Fil: Sarli, Macarena. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Sarli, Macarena. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: De Salvo, M.N. Instituto de Zoonosis Luis Pasteur; Argentina
Fil: Díaz Pérez, Paula M. Instituto de Zoonosis Luis Pasteur; Argentina
Fil: Cicuttin, Gabriel L. Instituto de Zoonosis Luis Pasteur; Argentina
Fil: Nava, Santiago. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Nava, Santiago. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Sebastian, Patrick. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Sebastian, Patrick. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
description The aim of this work was to develop a real-time PCR assay with a TaqMan® probe that detects a species-specific part of the 16S rDNA gene of Ehrlichia canis. Canine blood samples (n = 207), collected and tested by a conventional PCR assay within a study conducted by De Salvo et al., were simultaneously analyzed with the novel designed real-time PCR, and the results of both assays were compared. The agreement between the two methods was 97.6 % with a kappa value of 0.92186. Hereby, the standard error was 0.034416 and the 95 % confidence interval from 0.8544 to 0.98931. While the conventional PCR assay showed false negative results (2.42 %; 5/207), the real-time PCR assays showed a specificity of 100 %. The results of the current study showed that the developed assay presents sensitivity and specificity for the detection of E. canis in blood samples, adding a new tool for the diagnosis of this pathogen.
publishDate 2024
dc.date.none.fl_str_mv 2024-09-24T10:32:37Z
2024-09-24T10:32:37Z
2024-12
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/20.500.12123/19510
https://www.sciencedirect.com/science/article/pii/S0732889324003432
0732-8893
1879-0070
https://doi.org/10.1016/j.diagmicrobio.2024.116517
url http://hdl.handle.net/20.500.12123/19510
https://www.sciencedirect.com/science/article/pii/S0732889324003432
https://doi.org/10.1016/j.diagmicrobio.2024.116517
identifier_str_mv 0732-8893
1879-0070
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repograntAgreement/INTA/2019-PE-E5-I109-001, Convocatoria: Estudios para el control de enfermedades subtropicales y/o transmitidas por vectores (Tristeza Bovina, Garrapatas, Miasis, Tripanosomiasis, Lengua Azul y la
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 Diagnostic Microbiology and Infectious Disease 110 (4) : 116517 (December 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
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