Double antigen sandwich ELISA for detection of antibodies against Babesia bovis

Autores
Primo, María Evangelina; Valentini, Beatriz Susana; Mazzucco Panizza, Matilde Nahimé; Morel, Nicolas; Thompson, Carolina Soledad
Año de publicación
2026
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Babesia bovis is a hemoparasite responsible for bovine babesiosis, an economically important tickborne disease widespread in tropical and subtropical regions of the world. Serological tests are used in calves to evaluate the need for vaccination and the immunity achieved after vaccination. We developed, validated, and evaluated under field conditions a double-antigen sandwich ELISA (dasELISA) using a recombinant major surface antigen 2c (MSA2c) from a pathogenic and an attenuated B. bovis strain. The tests were validated using a panel of 1057 serum samples of known status (217 known-positive and 840 known-negative). Samples from 116 herds (n = 2771) were evaluated by indirect ELISA (iELISA) and dasELISA. Samples with disagreement between ELISAs (231 out of 594, 38.9 %) were analyzed by IFAT. The dasELISA cutoff was ≥ 52 %P, with a diagnostic sensitivity of 94.5 % (95 % CI, 90.5–97.1) and a diagnostic specificity of 96.4 % (95 % CI, 94.9–97.6). The area under the curve (AUC) was assessed to be 0.979, which corresponds to an excellent ability of the assay to discriminate truly infected from truly uninfected animals. Concordance among dasELISA and iELISA was 78.6 %, with a κ value of 0.567 (95 % CI = 0.537–0.598); meanwhile, 57 % and 43 % of the IFAT results agreed with dasELISA and iELISA, respectively. The dasELISA could be useful for large-scale detection of anti–B. bovis antibodies in calves pre- and post-vaccination and for seroepidemiologic investigations, given its appropriate sensitivity and specificity and the simplicity of production.
EEA Rafaela
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
Fil: Valentini, Beatriz. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina
Fil: Valentini, Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina
Fil: Mazzucco Panizza, Matilde Nahimé. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina
Fil: Mazzucco Panizza, Matilde Nahimé. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina
Fil: Morel, Nicolas. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina
Fil: Morel, Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina
Fil: Thompson, Carolina Soledad. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina
Fil: Thompson, Carolina Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina
Fuente
Veterinary Parasitology 342 : 110657. (February 2026)
Materia
Babesia bovis
Antigens
ELISA
Antibodies
Tick-borne Diseases
Cattle
Antígenos
Anticuerpos
Enfermedad Transmitida por Garrapatas
Ganado Bovino
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:20.500.12123/24883
Acceder
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oai_identifier_str oai:localhost:20.500.12123/24883
network_acronym_str INTADig
repository_id_str l
network_name_str INTA Digital (INTA)
spelling Double antigen sandwich ELISA for detection of antibodies against Babesia bovisPrimo, María EvangelinaValentini, Beatriz SusanaMazzucco Panizza, Matilde NahiméMorel, NicolasThompson, Carolina SoledadBabesia bovisAntigensELISAAntibodiesTick-borne DiseasesCattleAntígenosAnticuerposEnfermedad Transmitida por GarrapatasGanado BovinoBabesia bovis is a hemoparasite responsible for bovine babesiosis, an economically important tickborne disease widespread in tropical and subtropical regions of the world. Serological tests are used in calves to evaluate the need for vaccination and the immunity achieved after vaccination. We developed, validated, and evaluated under field conditions a double-antigen sandwich ELISA (dasELISA) using a recombinant major surface antigen 2c (MSA2c) from a pathogenic and an attenuated B. bovis strain. The tests were validated using a panel of 1057 serum samples of known status (217 known-positive and 840 known-negative). Samples from 116 herds (n = 2771) were evaluated by indirect ELISA (iELISA) and dasELISA. Samples with disagreement between ELISAs (231 out of 594, 38.9 %) were analyzed by IFAT. The dasELISA cutoff was ≥ 52 %P, with a diagnostic sensitivity of 94.5 % (95 % CI, 90.5–97.1) and a diagnostic specificity of 96.4 % (95 % CI, 94.9–97.6). The area under the curve (AUC) was assessed to be 0.979, which corresponds to an excellent ability of the assay to discriminate truly infected from truly uninfected animals. Concordance among dasELISA and iELISA was 78.6 %, with a κ value of 0.567 (95 % CI = 0.537–0.598); meanwhile, 57 % and 43 % of the IFAT results agreed with dasELISA and iELISA, respectively. The dasELISA could be useful for large-scale detection of anti–B. bovis antibodies in calves pre- and post-vaccination and for seroepidemiologic investigations, given its appropriate sensitivity and specificity and the simplicity of production.EEA RafaelaFil: 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); ArgentinaFil: Valentini, Beatriz. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); ArgentinaFil: Valentini, Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IdICaL); ArgentinaFil: Mazzucco Panizza, Matilde Nahimé. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); ArgentinaFil: Mazzucco Panizza, Matilde Nahimé. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IdICaL); ArgentinaFil: Morel, Nicolas. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); ArgentinaFil: Morel, Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IdICaL); ArgentinaFil: Thompson, Carolina Soledad. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); ArgentinaFil: Thompson, Carolina Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IdICaL); ArgentinaElsevier2026-01-05T16:32:46Z2026-01-05T16:32:46Z2026-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/24883https://www.sciencedirect.com/science/article/abs/pii/S03044017250026870304-40171873-2550https://doi.org/10.1016/j.vetpar.2025.110657Veterinary Parasitology 342 : 110657. (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)2026-01-08T10:41:02Zoai:localhost:20.500.12123/24883instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2026-01-08 10:41:02.926INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Double antigen sandwich ELISA for detection of antibodies against Babesia bovis
title Double antigen sandwich ELISA for detection of antibodies against Babesia bovis
spellingShingle Double antigen sandwich ELISA for detection of antibodies against Babesia bovis
Primo, María Evangelina
Babesia bovis
Antigens
ELISA
Antibodies
Tick-borne Diseases
Cattle
Antígenos
Anticuerpos
Enfermedad Transmitida por Garrapatas
Ganado Bovino
title_short Double antigen sandwich ELISA for detection of antibodies against Babesia bovis
title_full Double antigen sandwich ELISA for detection of antibodies against Babesia bovis
title_fullStr Double antigen sandwich ELISA for detection of antibodies against Babesia bovis
title_full_unstemmed Double antigen sandwich ELISA for detection of antibodies against Babesia bovis
title_sort Double antigen sandwich ELISA for detection of antibodies against Babesia bovis
dc.creator.none.fl_str_mv Primo, María Evangelina
Valentini, Beatriz Susana
Mazzucco Panizza, Matilde Nahimé
Morel, Nicolas
Thompson, Carolina Soledad
author Primo, María Evangelina
author_facet Primo, María Evangelina
Valentini, Beatriz Susana
Mazzucco Panizza, Matilde Nahimé
Morel, Nicolas
Thompson, Carolina Soledad
author_role author
author2 Valentini, Beatriz Susana
Mazzucco Panizza, Matilde Nahimé
Morel, Nicolas
Thompson, Carolina Soledad
author2_role author
author
author
author
dc.subject.none.fl_str_mv Babesia bovis
Antigens
ELISA
Antibodies
Tick-borne Diseases
Cattle
Antígenos
Anticuerpos
Enfermedad Transmitida por Garrapatas
Ganado Bovino
topic Babesia bovis
Antigens
ELISA
Antibodies
Tick-borne Diseases
Cattle
Antígenos
Anticuerpos
Enfermedad Transmitida por Garrapatas
Ganado Bovino
dc.description.none.fl_txt_mv Babesia bovis is a hemoparasite responsible for bovine babesiosis, an economically important tickborne disease widespread in tropical and subtropical regions of the world. Serological tests are used in calves to evaluate the need for vaccination and the immunity achieved after vaccination. We developed, validated, and evaluated under field conditions a double-antigen sandwich ELISA (dasELISA) using a recombinant major surface antigen 2c (MSA2c) from a pathogenic and an attenuated B. bovis strain. The tests were validated using a panel of 1057 serum samples of known status (217 known-positive and 840 known-negative). Samples from 116 herds (n = 2771) were evaluated by indirect ELISA (iELISA) and dasELISA. Samples with disagreement between ELISAs (231 out of 594, 38.9 %) were analyzed by IFAT. The dasELISA cutoff was ≥ 52 %P, with a diagnostic sensitivity of 94.5 % (95 % CI, 90.5–97.1) and a diagnostic specificity of 96.4 % (95 % CI, 94.9–97.6). The area under the curve (AUC) was assessed to be 0.979, which corresponds to an excellent ability of the assay to discriminate truly infected from truly uninfected animals. Concordance among dasELISA and iELISA was 78.6 %, with a κ value of 0.567 (95 % CI = 0.537–0.598); meanwhile, 57 % and 43 % of the IFAT results agreed with dasELISA and iELISA, respectively. The dasELISA could be useful for large-scale detection of anti–B. bovis antibodies in calves pre- and post-vaccination and for seroepidemiologic investigations, given its appropriate sensitivity and specificity and the simplicity of production.
EEA Rafaela
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
Fil: Valentini, Beatriz. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina
Fil: Valentini, Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina
Fil: Mazzucco Panizza, Matilde Nahimé. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina
Fil: Mazzucco Panizza, Matilde Nahimé. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina
Fil: Morel, Nicolas. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina
Fil: Morel, Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina
Fil: Thompson, Carolina Soledad. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina
Fil: Thompson, Carolina Soledad. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina
description Babesia bovis is a hemoparasite responsible for bovine babesiosis, an economically important tickborne disease widespread in tropical and subtropical regions of the world. Serological tests are used in calves to evaluate the need for vaccination and the immunity achieved after vaccination. We developed, validated, and evaluated under field conditions a double-antigen sandwich ELISA (dasELISA) using a recombinant major surface antigen 2c (MSA2c) from a pathogenic and an attenuated B. bovis strain. The tests were validated using a panel of 1057 serum samples of known status (217 known-positive and 840 known-negative). Samples from 116 herds (n = 2771) were evaluated by indirect ELISA (iELISA) and dasELISA. Samples with disagreement between ELISAs (231 out of 594, 38.9 %) were analyzed by IFAT. The dasELISA cutoff was ≥ 52 %P, with a diagnostic sensitivity of 94.5 % (95 % CI, 90.5–97.1) and a diagnostic specificity of 96.4 % (95 % CI, 94.9–97.6). The area under the curve (AUC) was assessed to be 0.979, which corresponds to an excellent ability of the assay to discriminate truly infected from truly uninfected animals. Concordance among dasELISA and iELISA was 78.6 %, with a κ value of 0.567 (95 % CI = 0.537–0.598); meanwhile, 57 % and 43 % of the IFAT results agreed with dasELISA and iELISA, respectively. The dasELISA could be useful for large-scale detection of anti–B. bovis antibodies in calves pre- and post-vaccination and for seroepidemiologic investigations, given its appropriate sensitivity and specificity and the simplicity of production.
publishDate 2026
dc.date.none.fl_str_mv 2026-01-05T16:32:46Z
2026-01-05T16:32:46Z
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/20.500.12123/24883
https://www.sciencedirect.com/science/article/abs/pii/S0304401725002687
0304-4017
1873-2550
https://doi.org/10.1016/j.vetpar.2025.110657
url http://hdl.handle.net/20.500.12123/24883
https://www.sciencedirect.com/science/article/abs/pii/S0304401725002687
https://doi.org/10.1016/j.vetpar.2025.110657
identifier_str_mv 0304-4017
1873-2550
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/restrictedAccess
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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 Veterinary Parasitology 342 : 110657. (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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