Development and Validation of a Novel ELISA for the Specific Detection of Antibodies against Mycobacterium avium Subspecies paratuberculosis Based on a Chimeric Polyprotein
Bovine paratuberculosis (PTB) is caused by Mycobacterium avium subsp. paratuberculosis (MAP). The optimization of detection tests specific for MAP is crucial to improve PTB control. In this work, we aimed to develop and validate a diagnostic tool based on an ELISA to specifically detect anti-MAP ant...
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| Format: | Article |
| Language: | English |
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Wiley
2021-01-01
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| Series: | Veterinary Medicine International |
| Online Access: | http://dx.doi.org/10.1155/2021/7336848 |
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| author | Roberto Damián Moyano Magali Andrea Romero María Alejandra Colombatti Olivieri María Fiorella Alvarado Pinedo Gabriel Eduardo Traveria María Isabel Romano María Natalia Alonso |
| author_facet | Roberto Damián Moyano Magali Andrea Romero María Alejandra Colombatti Olivieri María Fiorella Alvarado Pinedo Gabriel Eduardo Traveria María Isabel Romano María Natalia Alonso |
| author_sort | Roberto Damián Moyano |
| collection | DOAJ |
| description | Bovine paratuberculosis (PTB) is caused by Mycobacterium avium subsp. paratuberculosis (MAP). The optimization of detection tests specific for MAP is crucial to improve PTB control. In this work, we aimed to develop and validate a diagnostic tool based on an ELISA to specifically detect anti-MAP antibodies from bovine serum samples. For that purpose, we designed a recombinant polyprotein containing four specific antigens from MAP and optimized the ELISA. The validation consisted of the assessment of 10 sera from PTB-infected and healthy bovines with different OD values. The diagnostic performance of the polyprotein-ELISA was evaluated by testing 130 bovine serum samples (47 healthy, 48 MAP-infected, and 35 M. bovis-infected bovines). The ELISA using the polyprotein yielded an area under the ROC curve (AUC) of 0.9912 (95% CI, 0.9758–1.007; P < 0.0001). Moreover, for this ELISA, the cut-off selected from the ROC curve based on the point with a sensitivity of 95.56% (95% CI, 0.8485–0.9946) and specificity of 97.92 (95% CI, 0.8893–0.9995) was 0.3328. Similar results were obtained with an ELISA using the commercial Paratuberculosis Protoplasmatic Antigen (PPA). However, the ELISA with the polyprotein antigen showed a better performance against sera from animals infected with Mycobacterium bovis compared to the ELISA with PPA: lower cross-reactivity (2.85% versus 25.71%). These results demonstrate a very low cross-reactivity of the polyprotein with antibodies present in serum samples from animals infected with M. bovis. The designed polyprotein and the validated ELISA could be very useful for the specific identification of MAP-infected animals in herds. |
| format | Article |
| id | doaj-art-71e628d6b6eb4114adfe86a920e32517 |
| institution | Kabale University |
| issn | 2042-0048 |
| language | English |
| publishDate | 2021-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Veterinary Medicine International |
| spelling | doaj-art-71e628d6b6eb4114adfe86a920e325172025-02-03T05:58:23ZengWileyVeterinary Medicine International2042-00482021-01-01202110.1155/2021/7336848Development and Validation of a Novel ELISA for the Specific Detection of Antibodies against Mycobacterium avium Subspecies paratuberculosis Based on a Chimeric PolyproteinRoberto Damián Moyano0Magali Andrea Romero1María Alejandra Colombatti Olivieri2María Fiorella Alvarado Pinedo3Gabriel Eduardo Traveria4María Isabel Romano5María Natalia Alonso6Instituto de BiotecnologíaConsejo Nacional de Investigaciones Científicas y Técnicas (CONICET)Instituto de BiotecnologíaCentro de Diagnóstico e Investigaciones Veterinarias (CEDIVE) de la Facultad de Ciencias VeterinariasCentro de Diagnóstico e Investigaciones Veterinarias (CEDIVE) de la Facultad de Ciencias VeterinariasInstituto de BiotecnologíaInstituto de BiotecnologíaBovine paratuberculosis (PTB) is caused by Mycobacterium avium subsp. paratuberculosis (MAP). The optimization of detection tests specific for MAP is crucial to improve PTB control. In this work, we aimed to develop and validate a diagnostic tool based on an ELISA to specifically detect anti-MAP antibodies from bovine serum samples. For that purpose, we designed a recombinant polyprotein containing four specific antigens from MAP and optimized the ELISA. The validation consisted of the assessment of 10 sera from PTB-infected and healthy bovines with different OD values. The diagnostic performance of the polyprotein-ELISA was evaluated by testing 130 bovine serum samples (47 healthy, 48 MAP-infected, and 35 M. bovis-infected bovines). The ELISA using the polyprotein yielded an area under the ROC curve (AUC) of 0.9912 (95% CI, 0.9758–1.007; P < 0.0001). Moreover, for this ELISA, the cut-off selected from the ROC curve based on the point with a sensitivity of 95.56% (95% CI, 0.8485–0.9946) and specificity of 97.92 (95% CI, 0.8893–0.9995) was 0.3328. Similar results were obtained with an ELISA using the commercial Paratuberculosis Protoplasmatic Antigen (PPA). However, the ELISA with the polyprotein antigen showed a better performance against sera from animals infected with Mycobacterium bovis compared to the ELISA with PPA: lower cross-reactivity (2.85% versus 25.71%). These results demonstrate a very low cross-reactivity of the polyprotein with antibodies present in serum samples from animals infected with M. bovis. The designed polyprotein and the validated ELISA could be very useful for the specific identification of MAP-infected animals in herds.http://dx.doi.org/10.1155/2021/7336848 |
| spellingShingle | Roberto Damián Moyano Magali Andrea Romero María Alejandra Colombatti Olivieri María Fiorella Alvarado Pinedo Gabriel Eduardo Traveria María Isabel Romano María Natalia Alonso Development and Validation of a Novel ELISA for the Specific Detection of Antibodies against Mycobacterium avium Subspecies paratuberculosis Based on a Chimeric Polyprotein Veterinary Medicine International |
| title | Development and Validation of a Novel ELISA for the Specific Detection of Antibodies against Mycobacterium avium Subspecies paratuberculosis Based on a Chimeric Polyprotein |
| title_full | Development and Validation of a Novel ELISA for the Specific Detection of Antibodies against Mycobacterium avium Subspecies paratuberculosis Based on a Chimeric Polyprotein |
| title_fullStr | Development and Validation of a Novel ELISA for the Specific Detection of Antibodies against Mycobacterium avium Subspecies paratuberculosis Based on a Chimeric Polyprotein |
| title_full_unstemmed | Development and Validation of a Novel ELISA for the Specific Detection of Antibodies against Mycobacterium avium Subspecies paratuberculosis Based on a Chimeric Polyprotein |
| title_short | Development and Validation of a Novel ELISA for the Specific Detection of Antibodies against Mycobacterium avium Subspecies paratuberculosis Based on a Chimeric Polyprotein |
| title_sort | development and validation of a novel elisa for the specific detection of antibodies against mycobacterium avium subspecies paratuberculosis based on a chimeric polyprotein |
| url | http://dx.doi.org/10.1155/2021/7336848 |
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