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

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 fro...

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Main Authors: Bellezze, Julio, Thompson, Carolina Soledad, Bosio, Anabela, Torioni, Susana Marta, Primo, María Evangelina
Format: info:ar-repo/semantics/artículo
Language:Inglés
Published: American Association of Veterinary Laboratory Diagnosticians 2023
Subjects:
Online Access:http://hdl.handle.net/20.500.12123/14583
https://journals.sagepub.com/doi/10.1177/10406387231152472
https://doi.org/10.1177/10406387231152472
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author Bellezze, Julio
Thompson, Carolina Soledad
Bosio, Anabela
Torioni, Susana Marta
Primo, María Evangelina
author_browse Bellezze, Julio
Bosio, Anabela
Primo, María Evangelina
Thompson, Carolina Soledad
Torioni, Susana Marta
author_facet Bellezze, Julio
Thompson, Carolina Soledad
Bosio, Anabela
Torioni, Susana Marta
Primo, María Evangelina
author_sort Bellezze, Julio
collection INTA Digital
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.
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spelling INTA145832023-04-26T17:05:10Z Development and field evaluation of an ELISA to differentiate Anaplasma marginale–infected from A. centrale–vaccinated cattle Bellezze, Julio Thompson, Carolina Soledad Bosio, Anabela Torioni, Susana Marta Primo, María Evangelina Anaplasma marginale Anaplasma centrale ELISA Infectious Diseases Cattle Enfermedades Infecciosas Anaplasmosis Ganado Bovino Species-specific Antibodies Anticuerpos Específicos de Especies 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 2023-04-26T16:53:08Z 2023-04-26T16:53:08Z 2023-03 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion 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 eng 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) application/pdf American Association of Veterinary Laboratory Diagnosticians Journal of Veterinary Diagnostic Investigation 35 (2) : 204-208 (March 2023)
spellingShingle Anaplasma marginale
Anaplasma centrale
ELISA
Infectious Diseases
Cattle
Enfermedades Infecciosas
Anaplasmosis
Ganado Bovino
Species-specific Antibodies
Anticuerpos Específicos de Especies
Bellezze, Julio
Thompson, Carolina Soledad
Bosio, Anabela
Torioni, Susana Marta
Primo, María Evangelina
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
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_short 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
topic Anaplasma marginale
Anaplasma centrale
ELISA
Infectious Diseases
Cattle
Enfermedades Infecciosas
Anaplasmosis
Ganado Bovino
Species-specific Antibodies
Anticuerpos Específicos de Especies
url http://hdl.handle.net/20.500.12123/14583
https://journals.sagepub.com/doi/10.1177/10406387231152472
https://doi.org/10.1177/10406387231152472
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