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
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/159931
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/159931
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling 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 Técnicas2025-09-17T11:44:16Zoai: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:34982025-09-17 11:44:16.361CONICET 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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score 13.001348

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