Development and validation of a Novel ELISA for the specific detection of antibodies against Mycobacterium avium subspecies paratuberculosis based on a chimeric polyprotein
- Autores
- Moyano, Roberto Damian; Romero, Magali Andrea; Colombatti Olivieri, María Alejandra; Alvarado Pinedo, María Fiorella; Travería, Gabriel Eduardo; Romano, Maria Isabel; Alonso, Maria Natalia
- Año de publicación
- 2021
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- 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.
Fil: Moyano, Roberto Damian. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Romero, Magali Andrea. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Clínica. Centro de Diagnóstico e Investigaciones Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina
Fil: Colombatti Olivieri, María Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Alvarado Pinedo, María Fiorella. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Clínica. Centro de Diagnóstico e Investigaciones Veterinarias; Argentina
Fil: Travería, Gabriel Eduardo. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Clínica. Centro de Diagnóstico e Investigaciones Veterinarias; Argentina
Fil: Romano, Maria Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Alonso, Maria Natalia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina - Materia
-
Bovine paratuberculosis
ELISA
Chimeric polyprotein
Diagnosis - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/159931
Ver los metadatos del registro completo
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Development and validation of a Novel ELISA for the specific detection of antibodies against Mycobacterium avium subspecies paratuberculosis based on a chimeric polyproteinMoyano, Roberto DamianRomero, Magali AndreaColombatti Olivieri, María AlejandraAlvarado Pinedo, María FiorellaTravería, Gabriel EduardoRomano, Maria IsabelAlonso, Maria NataliaBovine paratuberculosisELISAChimeric polyproteinDiagnosishttps://purl.org/becyt/ford/4.3https://purl.org/becyt/ford/4Bovine 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.Fil: Moyano, Roberto Damian. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaFil: Romero, Magali Andrea. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Clínica. Centro de Diagnóstico e Investigaciones Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; ArgentinaFil: Colombatti Olivieri, María Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaFil: Alvarado Pinedo, María Fiorella. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Clínica. Centro de Diagnóstico e Investigaciones Veterinarias; ArgentinaFil: Travería, Gabriel Eduardo. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Clínica. Centro de Diagnóstico e Investigaciones Veterinarias; ArgentinaFil: Romano, Maria Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaFil: Alonso, Maria Natalia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; ArgentinaHindawi Publishing Corporation2021-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/159931Moyano, Roberto Damian; Romero, Magali Andrea; Colombatti Olivieri, María Alejandra; Alvarado Pinedo, María Fiorella; Travería, Gabriel Eduardo; et al.; Development and validation of a Novel ELISA for the specific detection of antibodies against Mycobacterium avium subspecies paratuberculosis based on a chimeric polyprotein; Hindawi Publishing Corporation; Veterinary Medicine International; 2021; 12-2021; 1-102042-0048CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1155/2021/7336848info:eu-repo/semantics/altIdentifier/url/https://www.hindawi.com/journals/vmi/2021/7336848/info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:07:35Zoai:ri.conicet.gov.ar:11336/159931instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 10:07:35.588CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
Development and validation of a Novel ELISA for the specific detection of antibodies against Mycobacterium avium subspecies paratuberculosis based on a chimeric polyprotein |
title |
Development and validation of a Novel ELISA for the specific detection of antibodies against Mycobacterium avium subspecies paratuberculosis based on a chimeric polyprotein |
spellingShingle |
Development and validation of a Novel ELISA for the specific detection of antibodies against Mycobacterium avium subspecies paratuberculosis based on a chimeric polyprotein Moyano, Roberto Damian Bovine paratuberculosis ELISA Chimeric polyprotein Diagnosis |
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_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_sort |
Development and validation of a Novel ELISA for the specific detection of antibodies against Mycobacterium avium subspecies paratuberculosis based on a chimeric polyprotein |
dc.creator.none.fl_str_mv |
Moyano, Roberto Damian Romero, Magali Andrea Colombatti Olivieri, María Alejandra Alvarado Pinedo, María Fiorella Travería, Gabriel Eduardo Romano, Maria Isabel Alonso, Maria Natalia |
author |
Moyano, Roberto Damian |
author_facet |
Moyano, Roberto Damian Romero, Magali Andrea Colombatti Olivieri, María Alejandra Alvarado Pinedo, María Fiorella Travería, Gabriel Eduardo Romano, Maria Isabel Alonso, Maria Natalia |
author_role |
author |
author2 |
Romero, Magali Andrea Colombatti Olivieri, María Alejandra Alvarado Pinedo, María Fiorella Travería, Gabriel Eduardo Romano, Maria Isabel Alonso, Maria Natalia |
author2_role |
author author author author author author |
dc.subject.none.fl_str_mv |
Bovine paratuberculosis ELISA Chimeric polyprotein Diagnosis |
topic |
Bovine paratuberculosis ELISA Chimeric polyprotein Diagnosis |
purl_subject.fl_str_mv |
https://purl.org/becyt/ford/4.3 https://purl.org/becyt/ford/4 |
dc.description.none.fl_txt_mv |
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. Fil: Moyano, Roberto Damian. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina Fil: Romero, Magali Andrea. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Clínica. Centro de Diagnóstico e Investigaciones Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina Fil: Colombatti Olivieri, María Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina Fil: Alvarado Pinedo, María Fiorella. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Clínica. Centro de Diagnóstico e Investigaciones Veterinarias; Argentina Fil: Travería, Gabriel Eduardo. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Departamento de Clínica. Centro de Diagnóstico e Investigaciones Veterinarias; Argentina Fil: Romano, Maria Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina Fil: Alonso, Maria Natalia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; Argentina |
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. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-12 |
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/11336/159931 Moyano, Roberto Damian; Romero, Magali Andrea; Colombatti Olivieri, María Alejandra; Alvarado Pinedo, María Fiorella; Travería, Gabriel Eduardo; et al.; Development and validation of a Novel ELISA for the specific detection of antibodies against Mycobacterium avium subspecies paratuberculosis based on a chimeric polyprotein; Hindawi Publishing Corporation; Veterinary Medicine International; 2021; 12-2021; 1-10 2042-0048 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/159931 |
identifier_str_mv |
Moyano, Roberto Damian; Romero, Magali Andrea; Colombatti Olivieri, María Alejandra; Alvarado Pinedo, María Fiorella; Travería, Gabriel Eduardo; et al.; Development and validation of a Novel ELISA for the specific detection of antibodies against Mycobacterium avium subspecies paratuberculosis based on a chimeric polyprotein; Hindawi Publishing Corporation; Veterinary Medicine International; 2021; 12-2021; 1-10 2042-0048 CONICET Digital CONICET |
dc.language.none.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
info:eu-repo/semantics/altIdentifier/doi/10.1155/2021/7336848 info:eu-repo/semantics/altIdentifier/url/https://www.hindawi.com/journals/vmi/2021/7336848/ |
dc.rights.none.fl_str_mv |
info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by/2.5/ar/ |
eu_rights_str_mv |
openAccess |
rights_invalid_str_mv |
https://creativecommons.org/licenses/by/2.5/ar/ |
dc.format.none.fl_str_mv |
application/pdf application/pdf |
dc.publisher.none.fl_str_mv |
Hindawi Publishing Corporation |
publisher.none.fl_str_mv |
Hindawi Publishing Corporation |
dc.source.none.fl_str_mv |
reponame:CONICET Digital (CONICET) instname:Consejo Nacional de Investigaciones Científicas y Técnicas |
reponame_str |
CONICET Digital (CONICET) |
collection |
CONICET Digital (CONICET) |
instname_str |
Consejo Nacional de Investigaciones Científicas y Técnicas |
repository.name.fl_str_mv |
CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas |
repository.mail.fl_str_mv |
dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar |
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1842270008318623744 |
score |
13.13397 |