Development and field evaluation of an ELISA to differentiate Anaplasma marginale–infected from A. centrale–vaccinated cattle

Autores
Bellezze, Julio; Thompson, Carolina Soledad; Bosio, Anabela; Torioni, Susana Marta; Primo, María Evangelina
Año de publicación
2023
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Immunization of calves with Anaplasma centrale is used to prevent acute anaplasmosis caused by A. marginale. Natural and vaccine-acquired immunity is detected through serologic tests based primarily on A. marginale recombinant major surface protein 5 (MSP5m) because it has 91% identity with MSP5 from A. centrale (MSP5c). We developed a displacement, double-antigen, sandwich ELISA (ddasELISA) to detect antibodies against A. marginale or A. centrale. For ddasELISA validation, we analyzed serum samples positive for antibodies against Anaplasma spp. from cattle naturally infected with A. marginale (n = 300) or vaccinated with A. centrale (n = 255). Species-specific nested PCR (nPCR) assays were used to confirm infection. The optical density (OD) values obtained from antibodies directed at unique epitopes of A. marginale (ODAm) or A. centrale (ODAc) were used in the formula ODAm/ODAc. If the derived ratio was >0.38, the serum sample was considered positive for antibodies against A. marginale, with 98.9% sensitivity and 98.0% specificity. In a field evaluation, we analyzed 702 Anaplasma spp. antibody–positive serum samples from 34 herds by ddasELISA and nPCR; 571 were classified by ddasELISA as A. marginale–infected or A. centrale–vaccinated, with 84% agreement (κ = 0.70) between ddasELISA and nPCR. Our results indicate that ddasELISA could be used as a cost-effective alternative to molecular techniques to confirm infection with A. marginale in countries in which prevention is based on vaccination with A. centrale.
INTA. Asociación Cooperadora INTA Rafaela (TCP 426100), ASATEC (IO-2019-130). Agencia Nacional de Promoción Científica y Tecnológica (PICT2013-0369, PICT2019-01498).
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: 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: Torioni, Susana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Torioni, 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: 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
Journal of Veterinary Diagnostic Investigation 35 (2) : 204-208 (March 2023)
Materia
Anaplasma marginale
Anaplasma centrale
ELISA
Infectious Diseases
Cattle
Enfermedades Infecciosas
Anaplasmosis
Ganado Bovino
Species-specific Antibodies
Anticuerpos Específicos de Especies
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:20.500.12123/14583
Acceder
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oai_identifier_str oai:localhost:20.500.12123/14583
network_acronym_str INTADig
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network_name_str INTA Digital (INTA)
spelling Development and field evaluation of an ELISA to differentiate Anaplasma marginale–infected from A. centrale–vaccinated cattleBellezze, JulioThompson, Carolina SoledadBosio, AnabelaTorioni, Susana MartaPrimo, María EvangelinaAnaplasma marginaleAnaplasma centraleELISAInfectious DiseasesCattleEnfermedades InfecciosasAnaplasmosisGanado BovinoSpecies-specific AntibodiesAnticuerpos Específicos de EspeciesImmunization of calves with Anaplasma centrale is used to prevent acute anaplasmosis caused by A. marginale. Natural and vaccine-acquired immunity is detected through serologic tests based primarily on A. marginale recombinant major surface protein 5 (MSP5m) because it has 91% identity with MSP5 from A. centrale (MSP5c). We developed a displacement, double-antigen, sandwich ELISA (ddasELISA) to detect antibodies against A. marginale or A. centrale. For ddasELISA validation, we analyzed serum samples positive for antibodies against Anaplasma spp. from cattle naturally infected with A. marginale (n = 300) or vaccinated with A. centrale (n = 255). Species-specific nested PCR (nPCR) assays were used to confirm infection. The optical density (OD) values obtained from antibodies directed at unique epitopes of A. marginale (ODAm) or A. centrale (ODAc) were used in the formula ODAm/ODAc. If the derived ratio was >0.38, the serum sample was considered positive for antibodies against A. marginale, with 98.9% sensitivity and 98.0% specificity. In a field evaluation, we analyzed 702 Anaplasma spp. antibody–positive serum samples from 34 herds by ddasELISA and nPCR; 571 were classified by ddasELISA as A. marginale–infected or A. centrale–vaccinated, with 84% agreement (κ = 0.70) between ddasELISA and nPCR. Our results indicate that ddasELISA could be used as a cost-effective alternative to molecular techniques to confirm infection with A. marginale in countries in which prevention is based on vaccination with A. centrale.INTA. Asociación Cooperadora INTA Rafaela (TCP 426100), ASATEC (IO-2019-130). Agencia Nacional de Promoción Científica y Tecnológica (PICT2013-0369, PICT2019-01498).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: 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: Torioni, Susana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); ArgentinaFil: Torioni, 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: 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); ArgentinaAmerican Association of Veterinary Laboratory Diagnosticians2023-04-26T16:53:08Z2023-04-26T16:53:08Z2023-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/14583https://journals.sagepub.com/doi/10.1177/10406387231152472Bellezze J, Thompson CS, Bosio AS, Torioni SM, Primo ME. Development and field evaluation of an ELISA to differentiate Anaplasma marginale–infected from A. centrale–vaccinated cattle. Journal of Veterinary Diagnostic Investigation. 2023;35(2):204-208. doi:10.1177/104063872311524721943-4936 (electronic)1040-6387https://doi.org/10.1177/10406387231152472Journal of Veterinary Diagnostic Investigation 35 (2) : 204-208 (March 2023)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)2025-09-11T10:24:36Zoai:localhost:20.500.12123/14583instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-11 10:24:36.948INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv Development and field evaluation of an ELISA to differentiate Anaplasma marginale–infected from A. centrale–vaccinated cattle
title Development and field evaluation of an ELISA to differentiate Anaplasma marginale–infected from A. centrale–vaccinated cattle
spellingShingle Development and field evaluation of an ELISA to differentiate Anaplasma marginale–infected from A. centrale–vaccinated cattle
Bellezze, Julio
Anaplasma marginale
Anaplasma centrale
ELISA
Infectious Diseases
Cattle
Enfermedades Infecciosas
Anaplasmosis
Ganado Bovino
Species-specific Antibodies
Anticuerpos Específicos de Especies
title_short Development and field evaluation of an ELISA to differentiate Anaplasma marginale–infected from A. centrale–vaccinated cattle
title_full Development and field evaluation of an ELISA to differentiate Anaplasma marginale–infected from A. centrale–vaccinated cattle
title_fullStr Development and field evaluation of an ELISA to differentiate Anaplasma marginale–infected from A. centrale–vaccinated cattle
title_full_unstemmed Development and field evaluation of an ELISA to differentiate Anaplasma marginale–infected from A. centrale–vaccinated cattle
title_sort Development and field evaluation of an ELISA to differentiate Anaplasma marginale–infected from A. centrale–vaccinated cattle
dc.creator.none.fl_str_mv Bellezze, Julio
Thompson, Carolina Soledad
Bosio, Anabela
Torioni, Susana Marta
Primo, María Evangelina
author Bellezze, Julio
author_facet Bellezze, Julio
Thompson, Carolina Soledad
Bosio, Anabela
Torioni, Susana Marta
Primo, María Evangelina
author_role author
author2 Thompson, Carolina Soledad
Bosio, Anabela
Torioni, Susana Marta
Primo, María Evangelina
author2_role author
author
author
author
dc.subject.none.fl_str_mv Anaplasma marginale
Anaplasma centrale
ELISA
Infectious Diseases
Cattle
Enfermedades Infecciosas
Anaplasmosis
Ganado Bovino
Species-specific Antibodies
Anticuerpos Específicos de Especies
topic Anaplasma marginale
Anaplasma centrale
ELISA
Infectious Diseases
Cattle
Enfermedades Infecciosas
Anaplasmosis
Ganado Bovino
Species-specific Antibodies
Anticuerpos Específicos de Especies
dc.description.none.fl_txt_mv Immunization of calves with Anaplasma centrale is used to prevent acute anaplasmosis caused by A. marginale. Natural and vaccine-acquired immunity is detected through serologic tests based primarily on A. marginale recombinant major surface protein 5 (MSP5m) because it has 91% identity with MSP5 from A. centrale (MSP5c). We developed a displacement, double-antigen, sandwich ELISA (ddasELISA) to detect antibodies against A. marginale or A. centrale. For ddasELISA validation, we analyzed serum samples positive for antibodies against Anaplasma spp. from cattle naturally infected with A. marginale (n = 300) or vaccinated with A. centrale (n = 255). Species-specific nested PCR (nPCR) assays were used to confirm infection. The optical density (OD) values obtained from antibodies directed at unique epitopes of A. marginale (ODAm) or A. centrale (ODAc) were used in the formula ODAm/ODAc. If the derived ratio was >0.38, the serum sample was considered positive for antibodies against A. marginale, with 98.9% sensitivity and 98.0% specificity. In a field evaluation, we analyzed 702 Anaplasma spp. antibody–positive serum samples from 34 herds by ddasELISA and nPCR; 571 were classified by ddasELISA as A. marginale–infected or A. centrale–vaccinated, with 84% agreement (κ = 0.70) between ddasELISA and nPCR. Our results indicate that ddasELISA could be used as a cost-effective alternative to molecular techniques to confirm infection with A. marginale in countries in which prevention is based on vaccination with A. centrale.
INTA. Asociación Cooperadora INTA Rafaela (TCP 426100), ASATEC (IO-2019-130). Agencia Nacional de Promoción Científica y Tecnológica (PICT2013-0369, PICT2019-01498).
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: 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: Torioni, Susana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina
Fil: Torioni, 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: 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 Immunization of calves with Anaplasma centrale is used to prevent acute anaplasmosis caused by A. marginale. Natural and vaccine-acquired immunity is detected through serologic tests based primarily on A. marginale recombinant major surface protein 5 (MSP5m) because it has 91% identity with MSP5 from A. centrale (MSP5c). We developed a displacement, double-antigen, sandwich ELISA (ddasELISA) to detect antibodies against A. marginale or A. centrale. For ddasELISA validation, we analyzed serum samples positive for antibodies against Anaplasma spp. from cattle naturally infected with A. marginale (n = 300) or vaccinated with A. centrale (n = 255). Species-specific nested PCR (nPCR) assays were used to confirm infection. The optical density (OD) values obtained from antibodies directed at unique epitopes of A. marginale (ODAm) or A. centrale (ODAc) were used in the formula ODAm/ODAc. If the derived ratio was >0.38, the serum sample was considered positive for antibodies against A. marginale, with 98.9% sensitivity and 98.0% specificity. In a field evaluation, we analyzed 702 Anaplasma spp. antibody–positive serum samples from 34 herds by ddasELISA and nPCR; 571 were classified by ddasELISA as A. marginale–infected or A. centrale–vaccinated, with 84% agreement (κ = 0.70) between ddasELISA and nPCR. Our results indicate that ddasELISA could be used as a cost-effective alternative to molecular techniques to confirm infection with A. marginale in countries in which prevention is based on vaccination with A. centrale.
publishDate 2023
dc.date.none.fl_str_mv 2023-04-26T16:53:08Z
2023-04-26T16:53:08Z
2023-03
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/20.500.12123/14583
https://journals.sagepub.com/doi/10.1177/10406387231152472
Bellezze J, Thompson CS, Bosio AS, Torioni SM, Primo ME. Development and field evaluation of an ELISA to differentiate Anaplasma marginale–infected from A. centrale–vaccinated cattle. Journal of Veterinary Diagnostic Investigation. 2023;35(2):204-208. doi:10.1177/10406387231152472
1943-4936 (electronic)
1040-6387
https://doi.org/10.1177/10406387231152472
url http://hdl.handle.net/20.500.12123/14583
https://journals.sagepub.com/doi/10.1177/10406387231152472
https://doi.org/10.1177/10406387231152472
identifier_str_mv Bellezze J, Thompson CS, Bosio AS, Torioni SM, Primo ME. Development and field evaluation of an ELISA to differentiate Anaplasma marginale–infected from A. centrale–vaccinated cattle. Journal of Veterinary Diagnostic Investigation. 2023;35(2):204-208. doi:10.1177/10406387231152472
1943-4936 (electronic)
1040-6387
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 American Association of Veterinary Laboratory Diagnosticians
publisher.none.fl_str_mv American Association of Veterinary Laboratory Diagnosticians
dc.source.none.fl_str_mv Journal of Veterinary Diagnostic Investigation 35 (2) : 204-208 (March 2023)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
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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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