Development and Bayesian validation of a competitive inhibition ELISA for detection of antibodies against Brucella abortus in cattle
Bovine brucellosis is a zoonotic disease caused by Brucella abortus, responsible for abortions in cows. It is endemic in low- and middle-income countries, where the brucellosis control and eradication programs are based on compulsory vaccination, detection of infected cattle through serologic assays...
| Main Authors: | , , , , , , |
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| Format: | Artículo |
| Language: | Inglés |
| Published: |
Elsevier
2025
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| Subjects: | |
| Online Access: | http://hdl.handle.net/20.500.12123/22165 https://www.sciencedirect.com/science/article/abs/pii/S0001706X24002018 https://doi.org/10.1016/j.actatropica.2024.107319 |
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| author | Novoa, María Belén Aguirre, Nerina Valentini, Beatriz Susana Signorini Porchiett, Marcelo Lisandro Aznar, Maria Natalia Vanzini, Victor Rene Torioni, Susana Marta |
| author_browse | Aguirre, Nerina Aznar, Maria Natalia Novoa, María Belén Signorini Porchiett, Marcelo Lisandro Torioni, Susana Marta Valentini, Beatriz Susana Vanzini, Victor Rene |
| author_facet | Novoa, María Belén Aguirre, Nerina Valentini, Beatriz Susana Signorini Porchiett, Marcelo Lisandro Aznar, Maria Natalia Vanzini, Victor Rene Torioni, Susana Marta |
| author_sort | Novoa, María Belén |
| collection | INTA Digital |
| description | Bovine brucellosis is a zoonotic disease caused by Brucella abortus, responsible for abortions in cows. It is endemic in low- and middle-income countries, where the brucellosis control and eradication programs are based on compulsory vaccination, detection of infected cattle through serologic assays, and culling of infected animals at slaughterhouses. The development of high sensitivity and specificity, and low-cost serologic assays guarantee their implementation in countries where the disease is endemic. The aim of the present study was to develop and validate a competitive inhibition enzyme-linked immune assay (ciELISA) to detect anti-B. abortus antibodies in sera from cattle. The developed ciELISA was validated using 2833 serum samples from dairy and beef cattle. From these, 1515 sera were from uninfected cows that belonged to free of brucellosis herds and 1318 were from infected cows that belonged positive to brucellosis herds. Sera were analyzed with the developed ciELISA, the buffer plate antigen (BPA) test, and the complement fixation test (CFT). The brucellosis status of the herds was officially established according to the country legislation and consistent for at least 5 years and was defined for each cow using the CFT as gold standard. The cutoff for the ciELISA was calculated using a ROC curve and its sensitivity and specificity were analyzed using the Bayesian Latent Class Model (BLCM) approach. The agreement among tests was calculated using the kappa (κ) value. In addition, 15 calves were vaccinated with 3 × 1010 viable cells of B. abortus Strain 19 vaccine, and the dynamics of antibodies were measured by CFT, buffered plate antigen (BPA) test, and the developed ciELISA. The obtained cutoff for ciELISA was ≥ 47 percentage of inhibition (% I), at the BLCM approach the sensitivity was 99.01 % (95 % CI: 97.55–100) and the specificity 98.74 % (95 % CI: 97.68–99.8). The κ between the ciELISA and BPA was κ = 0.88 and between the ciELISA and CFT κ = 0.95. Antibodies against B. abortus were detected in all the vaccinated calves 7 days after vaccination (AV) by the three assays, at day 135 AV all the calves were negative to CFT (15/15), 93.3 % (14/15) to ciELISA and 73.3 % (11/15) to BPA, and at day 190 AV all the calves were negative to the three assays. The developed ciELISA showed a very good performance, could detect the majority of vaccinated animals as negative after 135 days and could be used for the detection of anti-B. abortus antibodies in serum samples for the brucellosis control and eradication program. |
| format | Artículo |
| id | INTA22165 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2025 |
| publishDateRange | 2025 |
| publishDateSort | 2025 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | INTA221652025-05-06T11:16:40Z Development and Bayesian validation of a competitive inhibition ELISA for detection of antibodies against Brucella abortus in cattle Novoa, María Belén Aguirre, Nerina Valentini, Beatriz Susana Signorini Porchiett, Marcelo Lisandro Aznar, Maria Natalia Vanzini, Victor Rene Torioni, Susana Marta Ganado Bovino Enfermedades de los Animales Brucelosis Anticuerpos Cattle Animal Diseases Brucella abortus Brucellosis Antibodies ELISA Bovine brucellosis is a zoonotic disease caused by Brucella abortus, responsible for abortions in cows. It is endemic in low- and middle-income countries, where the brucellosis control and eradication programs are based on compulsory vaccination, detection of infected cattle through serologic assays, and culling of infected animals at slaughterhouses. The development of high sensitivity and specificity, and low-cost serologic assays guarantee their implementation in countries where the disease is endemic. The aim of the present study was to develop and validate a competitive inhibition enzyme-linked immune assay (ciELISA) to detect anti-B. abortus antibodies in sera from cattle. The developed ciELISA was validated using 2833 serum samples from dairy and beef cattle. From these, 1515 sera were from uninfected cows that belonged to free of brucellosis herds and 1318 were from infected cows that belonged positive to brucellosis herds. Sera were analyzed with the developed ciELISA, the buffer plate antigen (BPA) test, and the complement fixation test (CFT). The brucellosis status of the herds was officially established according to the country legislation and consistent for at least 5 years and was defined for each cow using the CFT as gold standard. The cutoff for the ciELISA was calculated using a ROC curve and its sensitivity and specificity were analyzed using the Bayesian Latent Class Model (BLCM) approach. The agreement among tests was calculated using the kappa (κ) value. In addition, 15 calves were vaccinated with 3 × 1010 viable cells of B. abortus Strain 19 vaccine, and the dynamics of antibodies were measured by CFT, buffered plate antigen (BPA) test, and the developed ciELISA. The obtained cutoff for ciELISA was ≥ 47 percentage of inhibition (% I), at the BLCM approach the sensitivity was 99.01 % (95 % CI: 97.55–100) and the specificity 98.74 % (95 % CI: 97.68–99.8). The κ between the ciELISA and BPA was κ = 0.88 and between the ciELISA and CFT κ = 0.95. Antibodies against B. abortus were detected in all the vaccinated calves 7 days after vaccination (AV) by the three assays, at day 135 AV all the calves were negative to CFT (15/15), 93.3 % (14/15) to ciELISA and 73.3 % (11/15) to BPA, and at day 190 AV all the calves were negative to the three assays. The developed ciELISA showed a very good performance, could detect the majority of vaccinated animals as negative after 135 days and could be used for the detection of anti-B. abortus antibodies in serum samples for the brucellosis control and eradication program. EEA Rafaela Fil: Novoa, María Belen. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina Fil: Novoa, María Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina Fil: Aguirre, Nerina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina Fil: Aguirre, Nerina. 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: Signorini, Marcelo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina Fil: Signorini, Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina Fil: Aznar, María Natalia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patobiología Veterinaria (IPVET); Argentina Fil: Vanzini, Victor Rene. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina Fil: Vanzini, Victor Rene. 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 2025-05-06T11:13:19Z 2025-05-06T11:13:19Z 2024-09 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/22165 https://www.sciencedirect.com/science/article/abs/pii/S0001706X24002018 0001-706X 1873-6254 https://doi.org/10.1016/j.actatropica.2024.107319 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 Elsevier Acta Tropica 257 : 107319. (September 2024) |
| spellingShingle | Ganado Bovino Enfermedades de los Animales Brucelosis Anticuerpos Cattle Animal Diseases Brucella abortus Brucellosis Antibodies ELISA Novoa, María Belén Aguirre, Nerina Valentini, Beatriz Susana Signorini Porchiett, Marcelo Lisandro Aznar, Maria Natalia Vanzini, Victor Rene Torioni, Susana Marta Development and Bayesian validation of a competitive inhibition ELISA for detection of antibodies against Brucella abortus in cattle |
| title | Development and Bayesian validation of a competitive inhibition ELISA for detection of antibodies against Brucella abortus in cattle |
| title_full | Development and Bayesian validation of a competitive inhibition ELISA for detection of antibodies against Brucella abortus in cattle |
| title_fullStr | Development and Bayesian validation of a competitive inhibition ELISA for detection of antibodies against Brucella abortus in cattle |
| title_full_unstemmed | Development and Bayesian validation of a competitive inhibition ELISA for detection of antibodies against Brucella abortus in cattle |
| title_short | Development and Bayesian validation of a competitive inhibition ELISA for detection of antibodies against Brucella abortus in cattle |
| title_sort | development and bayesian validation of a competitive inhibition elisa for detection of antibodies against brucella abortus in cattle |
| topic | Ganado Bovino Enfermedades de los Animales Brucelosis Anticuerpos Cattle Animal Diseases Brucella abortus Brucellosis Antibodies ELISA |
| url | http://hdl.handle.net/20.500.12123/22165 https://www.sciencedirect.com/science/article/abs/pii/S0001706X24002018 https://doi.org/10.1016/j.actatropica.2024.107319 |
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