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

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.

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Main Authors: Sarli, Macarena, De Salvo, María N., Díaz Pérez, Paula M., Cicuttin, Gabriel L., Nava, Santiago, Sebastian, Patrick
Format: info:ar-repo/semantics/artículo biblioteca
Language:eng
Published: Elsevier 2024-12
Subjects:Tick-borne Diseases, Diagnosis, Veterinary Medicine, Dogs, Enfermedad Transmitida por Garrapatas, Diagnóstico, Medicina Veterinaria, Ehrlichia canis, Perro, Hemoparasite, Hemoparásitos,
Online Access: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
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spelling oai:localhost:20.500.12123-195102024-09-24T10:53:24Z Development and evaluation of a TaqMan® real-time PCR assay for species-specific detection of Ehrlichia canis Sarli, Macarena De Salvo, María N. Díaz Pérez, Paula M. Cicuttin, Gabriel L. Nava, Santiago Sebastian, Patrick Tick-borne Diseases Diagnosis Veterinary Medicine Dogs Enfermedad Transmitida por Garrapatas Diagnóstico Medicina Veterinaria Ehrlichia canis Perro Hemoparasite Hemoparásitos 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 2024-09-24T10:32:37Z 2024-09-24T10:32:37Z 2024-12 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion 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 eng 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 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) application/pdf Elsevier Diagnostic Microbiology and Infectious Disease 110 (4) : 116517 (December 2024)
institution INTA AR
collection DSpace
country Argentina
countrycode AR
component Bibliográfico
access En linea
databasecode dig-inta-ar
tag biblioteca
region America del Sur
libraryname Biblioteca Central del INTA Argentina
language eng
topic Tick-borne Diseases
Diagnosis
Veterinary Medicine
Dogs
Enfermedad Transmitida por Garrapatas
Diagnóstico
Medicina Veterinaria
Ehrlichia canis
Perro
Hemoparasite
Hemoparásitos
Tick-borne Diseases
Diagnosis
Veterinary Medicine
Dogs
Enfermedad Transmitida por Garrapatas
Diagnóstico
Medicina Veterinaria
Ehrlichia canis
Perro
Hemoparasite
Hemoparásitos
spellingShingle Tick-borne Diseases
Diagnosis
Veterinary Medicine
Dogs
Enfermedad Transmitida por Garrapatas
Diagnóstico
Medicina Veterinaria
Ehrlichia canis
Perro
Hemoparasite
Hemoparásitos
Tick-borne Diseases
Diagnosis
Veterinary Medicine
Dogs
Enfermedad Transmitida por Garrapatas
Diagnóstico
Medicina Veterinaria
Ehrlichia canis
Perro
Hemoparasite
Hemoparásitos
Sarli, Macarena
De Salvo, María N.
Díaz Pérez, Paula M.
Cicuttin, Gabriel L.
Nava, Santiago
Sebastian, Patrick
Development and evaluation of a TaqMan® real-time PCR assay for species-specific detection of Ehrlichia canis
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.
format info:ar-repo/semantics/artículo
topic_facet Tick-borne Diseases
Diagnosis
Veterinary Medicine
Dogs
Enfermedad Transmitida por Garrapatas
Diagnóstico
Medicina Veterinaria
Ehrlichia canis
Perro
Hemoparasite
Hemoparásitos
author Sarli, Macarena
De Salvo, María N.
Díaz Pérez, Paula M.
Cicuttin, Gabriel L.
Nava, Santiago
Sebastian, Patrick
author_facet Sarli, Macarena
De Salvo, María N.
Díaz Pérez, Paula M.
Cicuttin, Gabriel L.
Nava, Santiago
Sebastian, Patrick
author_sort Sarli, Macarena
title Development and evaluation of a TaqMan® real-time PCR assay for species-specific detection of Ehrlichia canis
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
publisher Elsevier
publishDate 2024-12
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
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