Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR
The aim of this study was to standardize a diagnosis procedure to detect Mycobacterium avium subsp. paratuberculosis (Map) DNA in raw cow milk samples under field conditions. A procedure that combines both immunomagnetic separation and IS900-PCR detection (IMS-IS1 PCR) was employed on milk samples f...
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| Format: | info:ar-repo/semantics/artículo |
| Language: | Inglés |
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Elsevier
2019
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| Subjects: | |
| Online Access: | http://hdl.handle.net/20.500.12123/4944 https://www.sciencedirect.com/science/article/pii/S1517838216000605 https://doi.org/10.1016/j.bjm.2016.01.013 |
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| author | Gilardoni, Liliana Rosa Fernández, Bárbara Morsella, Claudia Graciela Mendez, Laura Luján Jar, Ana María Paolicchi, Fernando Mundo, Silvia Leonor |
| author_browse | Fernández, Bárbara Gilardoni, Liliana Rosa Jar, Ana María Mendez, Laura Luján Morsella, Claudia Graciela Mundo, Silvia Leonor Paolicchi, Fernando |
| author_facet | Gilardoni, Liliana Rosa Fernández, Bárbara Morsella, Claudia Graciela Mendez, Laura Luján Jar, Ana María Paolicchi, Fernando Mundo, Silvia Leonor |
| author_sort | Gilardoni, Liliana Rosa |
| collection | INTA Digital |
| description | The aim of this study was to standardize a diagnosis procedure to detect Mycobacterium avium subsp. paratuberculosis (Map) DNA in raw cow milk samples under field conditions. A procedure that combines both immunomagnetic separation and IS900-PCR detection (IMS-IS1 PCR) was employed on milk samples from 265 lactating Holstein cows from Map infected and uninfected herds in Argentina. IMS-IS1 PCR results were analyzed and compared with those obtained from milk and fecal culture and serum ELISA. The extent of agreement between both tests was determined by the Kappa test. IMS-IS1 PCR showed a detection limit of 101 CFU of Map/mL of milk, when 50:50 mix of monoclonal and polyclonal antibodies were used to coat magnetic beads. All of the 118 samples from the Map uninfected herds were negative for the set of the tests. In Map infected herds, 80 out of 147 cows tested positive by milk IMS-IS1 PCR (55%), of which 2 (1.4%) were also positive by milk culture, 15 (10%) by fecal culture, and 20 (14%) by serum ELISA. Kappa statistics (95% CI) showed a slight agreement between the different tests (<0.20), and the proportions of agreement were ≤0.55. The IMS-IS1 PCR method detected Map in milk of the cows that were not positive in other techniques. This is the first report dealing with the application of IMS-IS1 PCR in the detection of Map in raw milk samples under field conditions in Argentina. |
| format | info:ar-repo/semantics/artículo |
| id | INTA4944 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2019 |
| publishDateRange | 2019 |
| publishDateSort | 2019 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | INTA49442022-11-30T10:00:34Z Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR Gilardoni, Liliana Rosa Fernández, Bárbara Morsella, Claudia Graciela Mendez, Laura Luján Jar, Ana María Paolicchi, Fernando Mundo, Silvia Leonor Mycobacterium avium Leche de Vaca PCR Técnicas Inmunológicas Cow Milk Immunological Techniques Argentina Mycobacterium paratuberculosis Reacción en Cadena de la Polimerasa The aim of this study was to standardize a diagnosis procedure to detect Mycobacterium avium subsp. paratuberculosis (Map) DNA in raw cow milk samples under field conditions. A procedure that combines both immunomagnetic separation and IS900-PCR detection (IMS-IS1 PCR) was employed on milk samples from 265 lactating Holstein cows from Map infected and uninfected herds in Argentina. IMS-IS1 PCR results were analyzed and compared with those obtained from milk and fecal culture and serum ELISA. The extent of agreement between both tests was determined by the Kappa test. IMS-IS1 PCR showed a detection limit of 101 CFU of Map/mL of milk, when 50:50 mix of monoclonal and polyclonal antibodies were used to coat magnetic beads. All of the 118 samples from the Map uninfected herds were negative for the set of the tests. In Map infected herds, 80 out of 147 cows tested positive by milk IMS-IS1 PCR (55%), of which 2 (1.4%) were also positive by milk culture, 15 (10%) by fecal culture, and 20 (14%) by serum ELISA. Kappa statistics (95% CI) showed a slight agreement between the different tests (<0.20), and the proportions of agreement were ≤0.55. The IMS-IS1 PCR method detected Map in milk of the cows that were not positive in other techniques. This is the first report dealing with the application of IMS-IS1 PCR in the detection of Map in raw milk samples under field conditions in Argentina. EEA Balcarce Fil: Gilardoni, Liliana Rosa. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Cátedra de Inmunología; Argentina Fil: Fernández, Bárbara. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Cátedra de Inmunología; Argentina Fil: Morsella, Claudia Graciela. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce. Laboratorio de Bacteriología; Argentina Fil: Mendez, Laura Luján. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce. Laboratorio de Bacteriología; Argentina Fil: Jar, Ana María. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Cátedra de Inmunología; Argentina Fil: Paolicchi, Fernando. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce. Laboratorio de Bacteriología; Argentina Fil: Mundo, Silvia Leonor. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Cátedra de Inmunología; Argentina 2019-04-22T14:48:15Z 2019-04-22T14:48:15Z 2016-06 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/4944 https://www.sciencedirect.com/science/article/pii/S1517838216000605 1517-8382 1678-4405 https://doi.org/10.1016/j.bjm.2016.01.013 eng info:eu-repo/semantics/openAccess 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 Brazilian Journal of Microbiology 47 (2) : 506-512 (April–June 2016) |
| spellingShingle | Mycobacterium avium Leche de Vaca PCR Técnicas Inmunológicas Cow Milk Immunological Techniques Argentina Mycobacterium paratuberculosis Reacción en Cadena de la Polimerasa Gilardoni, Liliana Rosa Fernández, Bárbara Morsella, Claudia Graciela Mendez, Laura Luján Jar, Ana María Paolicchi, Fernando Mundo, Silvia Leonor Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR |
| title | Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR |
| title_full | Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR |
| title_fullStr | Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR |
| title_full_unstemmed | Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR |
| title_short | Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR |
| title_sort | mycobacterium paratuberculosis detection in cow s milk in argentina by immunomagnetic separation pcr |
| topic | Mycobacterium avium Leche de Vaca PCR Técnicas Inmunológicas Cow Milk Immunological Techniques Argentina Mycobacterium paratuberculosis Reacción en Cadena de la Polimerasa |
| url | http://hdl.handle.net/20.500.12123/4944 https://www.sciencedirect.com/science/article/pii/S1517838216000605 https://doi.org/10.1016/j.bjm.2016.01.013 |
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