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 simultaneous...
| Autores principales: | , , , , , |
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| Formato: | info:ar-repo/semantics/artículo |
| Lenguaje: | Inglés |
| Publicado: |
Elsevier
2024
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| Materias: | |
| Acceso en línea: | 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 |
| _version_ | 1855038025522741248 |
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| author | Sarli, Macarena De Salvo, María N. Díaz Pérez, Paula M. Cicuttin, Gabriel L. Nava, Santiago Sebastian, Patrick |
| author_browse | Cicuttin, Gabriel L. De Salvo, María N. Díaz Pérez, Paula M. Nava, Santiago Sarli, Macarena 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 |
| collection | INTA Digital |
| 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 |
| id | INTA19510 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2024 |
| publishDateRange | 2024 |
| publishDateSort | 2024 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | INTA195102024-09-24T10:53:24Z 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) |
| spellingShingle | 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 |
| title | 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_short | 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 r real time pcr assay for species specific detection of ehrlichia canis |
| topic | Tick-borne Diseases Diagnosis Veterinary Medicine Dogs Enfermedad Transmitida por Garrapatas Diagnóstico Medicina Veterinaria Ehrlichia canis Perro Hemoparasite Hemoparásitos |
| 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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