Development and field evaluation of a competitive ELISA to detect specific antibodies against Anaplasma marginale

Autores
Bellezze, Julio; Valentini, Beatriz Susana; Bosio, Anabela; Thompson, Carolina Soledad; Primo, María Evangelina
Año de publicación
2026
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
A species-specific competitive ELISA for Anaplasma marginale antibodies detection (cELISA-sAm) was developed by using both a monoclonal antibody specific to an epitope of MSP5 of A. marginale (Am6-mAb) and adding a pre-incubation phase with the truncated MSP5 of A. centrale (tMSP5c). For the determination of the cutoff value, diagnostic sensitivity (DSe) and specificity (DSp) of cELISA-sAm, 382 samples from cattle not infected with Anaplasma spp., 237 samples from cattle naturally infected with A. marginale, and 102 samples from cattle vaccinated with A. centrale were analyzed. The infection status was established using nested PCR (nPCR) or nPCR-restriction fragment length polymorphism assay (nPCR-RFLP) as reference technique. In a field evaluation, cELISA-sAm and nPCR-RFLP were used on Anaplasma spp. positive samples by double-antigen sandwich ELISA (dasELISA) from 2 herds to confirm the A. marginale infection. The cELISA-sAm optimal cutoff value was ≥ 18 %I, with a DSe of 89.9 % and a DSp of 99.0 %. In a field evaluation, concordance between cELISA-sAm and nPCR-RFLP was 93.0 % with κ = 0.81 (95 % CI = 0.56–1.00) for A. marginale-infected cattle detection. In countries in which prevention is based on vaccination with A. centrale, the replacement of current ELISAs by this cELISA-sAm would decrease the false positive results and avoid the need of species confirmation by molecular techniques when disease control programs are performed to monitor the prevalence of A. marginale.
EEA Rafaela
Fil: Bellezze, Julio. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Bellezze, Julio. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Valentini, Beatriz Susana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Valentini, Beatriz Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Bosio, Anabela. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Bosio, Anabela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Thompson, Carolina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Thompson, Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Primo, María Evangelina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Primo, María Evangelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fuente
The Veterinary Journal 315 : 106528. (February 2026)
Materia
Anaplasma marginale
ELISA
Antibodies
Cattle
Immune Response
Monoclonal Antibodies
Anticuerpos
Ganado Bovino
Respuesta Inmunológica
Anticuerpos Monoclonales
Nivel de accesibilidad
acceso restringido
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/24969
Acceder
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oai_identifier_str oai:localhost:20.500.12123/24969
network_acronym_str INTADig
repository_id_str l
network_name_str INTA Digital (INTA)
spelling Development and field evaluation of a competitive ELISA to detect specific antibodies against Anaplasma marginaleBellezze, JulioValentini, Beatriz SusanaBosio, AnabelaThompson, Carolina SoledadPrimo, María EvangelinaAnaplasma marginaleELISAAntibodiesCattleImmune ResponseMonoclonal AntibodiesAnticuerposGanado BovinoRespuesta InmunológicaAnticuerpos MonoclonalesA species-specific competitive ELISA for Anaplasma marginale antibodies detection (cELISA-sAm) was developed by using both a monoclonal antibody specific to an epitope of MSP5 of A. marginale (Am6-mAb) and adding a pre-incubation phase with the truncated MSP5 of A. centrale (tMSP5c). For the determination of the cutoff value, diagnostic sensitivity (DSe) and specificity (DSp) of cELISA-sAm, 382 samples from cattle not infected with Anaplasma spp., 237 samples from cattle naturally infected with A. marginale, and 102 samples from cattle vaccinated with A. centrale were analyzed. The infection status was established using nested PCR (nPCR) or nPCR-restriction fragment length polymorphism assay (nPCR-RFLP) as reference technique. In a field evaluation, cELISA-sAm and nPCR-RFLP were used on Anaplasma spp. positive samples by double-antigen sandwich ELISA (dasELISA) from 2 herds to confirm the A. marginale infection. The cELISA-sAm optimal cutoff value was ≥ 18 %I, with a DSe of 89.9 % and a DSp of 99.0 %. In a field evaluation, concordance between cELISA-sAm and nPCR-RFLP was 93.0 % with κ = 0.81 (95 % CI = 0.56–1.00) for A. marginale-infected cattle detection. In countries in which prevention is based on vaccination with A. centrale, the replacement of current ELISAs by this cELISA-sAm would decrease the false positive results and avoid the need of species confirmation by molecular techniques when disease control programs are performed to monitor the prevalence of A. marginale.EEA RafaelaFil: Bellezze, Julio. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); ArgentinaFil: Bellezze, Julio. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); ArgentinaFil: Valentini, Beatriz Susana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); ArgentinaFil: Valentini, Beatriz Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); ArgentinaFil: Bosio, Anabela. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); ArgentinaFil: Bosio, Anabela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); ArgentinaFil: Thompson, Carolina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); ArgentinaFil: Thompson, Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); ArgentinaFil: Primo, María Evangelina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); ArgentinaFil: Primo, María Evangelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); ArgentinaElsevier2026-01-09T14:46:19Z2026-01-09T14:46:19Z2026-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/24969https://www.sciencedirect.com/science/article/abs/pii/S10900233250023211090-02331532-2971https://doi.org/10.1016/j.tvjl.2025.106528The Veterinary Journal 315 : 106528. (February 2026)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo:eu-repo/semantics/restrictedAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2026-02-26T11:47:40Zoai:localhost:20.500.12123/24969instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2026-02-26 11:47:40.651INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Development and field evaluation of a competitive ELISA to detect specific antibodies against Anaplasma marginale
title Development and field evaluation of a competitive ELISA to detect specific antibodies against Anaplasma marginale
spellingShingle Development and field evaluation of a competitive ELISA to detect specific antibodies against Anaplasma marginale
Bellezze, Julio
Anaplasma marginale
ELISA
Antibodies
Cattle
Immune Response
Monoclonal Antibodies
Anticuerpos
Ganado Bovino
Respuesta Inmunológica
Anticuerpos Monoclonales
title_short Development and field evaluation of a competitive ELISA to detect specific antibodies against Anaplasma marginale
title_full Development and field evaluation of a competitive ELISA to detect specific antibodies against Anaplasma marginale
title_fullStr Development and field evaluation of a competitive ELISA to detect specific antibodies against Anaplasma marginale
title_full_unstemmed Development and field evaluation of a competitive ELISA to detect specific antibodies against Anaplasma marginale
title_sort Development and field evaluation of a competitive ELISA to detect specific antibodies against Anaplasma marginale
dc.creator.none.fl_str_mv Bellezze, Julio
Valentini, Beatriz Susana
Bosio, Anabela
Thompson, Carolina Soledad
Primo, María Evangelina
author Bellezze, Julio
author_facet Bellezze, Julio
Valentini, Beatriz Susana
Bosio, Anabela
Thompson, Carolina Soledad
Primo, María Evangelina
author_role author
author2 Valentini, Beatriz Susana
Bosio, Anabela
Thompson, Carolina Soledad
Primo, María Evangelina
author2_role author
author
author
author
dc.subject.none.fl_str_mv Anaplasma marginale
ELISA
Antibodies
Cattle
Immune Response
Monoclonal Antibodies
Anticuerpos
Ganado Bovino
Respuesta Inmunológica
Anticuerpos Monoclonales
topic Anaplasma marginale
ELISA
Antibodies
Cattle
Immune Response
Monoclonal Antibodies
Anticuerpos
Ganado Bovino
Respuesta Inmunológica
Anticuerpos Monoclonales
dc.description.none.fl_txt_mv A species-specific competitive ELISA for Anaplasma marginale antibodies detection (cELISA-sAm) was developed by using both a monoclonal antibody specific to an epitope of MSP5 of A. marginale (Am6-mAb) and adding a pre-incubation phase with the truncated MSP5 of A. centrale (tMSP5c). For the determination of the cutoff value, diagnostic sensitivity (DSe) and specificity (DSp) of cELISA-sAm, 382 samples from cattle not infected with Anaplasma spp., 237 samples from cattle naturally infected with A. marginale, and 102 samples from cattle vaccinated with A. centrale were analyzed. The infection status was established using nested PCR (nPCR) or nPCR-restriction fragment length polymorphism assay (nPCR-RFLP) as reference technique. In a field evaluation, cELISA-sAm and nPCR-RFLP were used on Anaplasma spp. positive samples by double-antigen sandwich ELISA (dasELISA) from 2 herds to confirm the A. marginale infection. The cELISA-sAm optimal cutoff value was ≥ 18 %I, with a DSe of 89.9 % and a DSp of 99.0 %. In a field evaluation, concordance between cELISA-sAm and nPCR-RFLP was 93.0 % with κ = 0.81 (95 % CI = 0.56–1.00) for A. marginale-infected cattle detection. In countries in which prevention is based on vaccination with A. centrale, the replacement of current ELISAs by this cELISA-sAm would decrease the false positive results and avoid the need of species confirmation by molecular techniques when disease control programs are performed to monitor the prevalence of A. marginale.
EEA Rafaela
Fil: Bellezze, Julio. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Bellezze, Julio. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Valentini, Beatriz Susana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Valentini, Beatriz Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Bosio, Anabela. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Bosio, Anabela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Thompson, Carolina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Thompson, Carolina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Primo, María Evangelina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Primo, María Evangelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
description A species-specific competitive ELISA for Anaplasma marginale antibodies detection (cELISA-sAm) was developed by using both a monoclonal antibody specific to an epitope of MSP5 of A. marginale (Am6-mAb) and adding a pre-incubation phase with the truncated MSP5 of A. centrale (tMSP5c). For the determination of the cutoff value, diagnostic sensitivity (DSe) and specificity (DSp) of cELISA-sAm, 382 samples from cattle not infected with Anaplasma spp., 237 samples from cattle naturally infected with A. marginale, and 102 samples from cattle vaccinated with A. centrale were analyzed. The infection status was established using nested PCR (nPCR) or nPCR-restriction fragment length polymorphism assay (nPCR-RFLP) as reference technique. In a field evaluation, cELISA-sAm and nPCR-RFLP were used on Anaplasma spp. positive samples by double-antigen sandwich ELISA (dasELISA) from 2 herds to confirm the A. marginale infection. The cELISA-sAm optimal cutoff value was ≥ 18 %I, with a DSe of 89.9 % and a DSp of 99.0 %. In a field evaluation, concordance between cELISA-sAm and nPCR-RFLP was 93.0 % with κ = 0.81 (95 % CI = 0.56–1.00) for A. marginale-infected cattle detection. In countries in which prevention is based on vaccination with A. centrale, the replacement of current ELISAs by this cELISA-sAm would decrease the false positive results and avoid the need of species confirmation by molecular techniques when disease control programs are performed to monitor the prevalence of A. marginale.
publishDate 2026
dc.date.none.fl_str_mv 2026-01-09T14:46:19Z
2026-01-09T14:46:19Z
2026-02
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/20.500.12123/24969
https://www.sciencedirect.com/science/article/abs/pii/S1090023325002321
1090-0233
1532-2971
https://doi.org/10.1016/j.tvjl.2025.106528
url http://hdl.handle.net/20.500.12123/24969
https://www.sciencedirect.com/science/article/abs/pii/S1090023325002321
https://doi.org/10.1016/j.tvjl.2025.106528
identifier_str_mv 1090-0233
1532-2971
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv 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)
eu_rights_str_mv restrictedAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv The Veterinary Journal 315 : 106528. (February 2026)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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