Development, validation and field evaluation of an indirect ELISA for the detection of antibodies against Brucella abortus in bulk and individual milk samples in dairy cattle
Brucellosis is an abortigenic and zoonotic disease. In cattle, it is mainly caused by Brucella abortus. The disease is endemic in low- and middle-income countries, being considered a neglected zoonotic disease. In these countries, it is of high importance to develop and validate sensitive, specific...
| Autores principales: | , , , , , , , |
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| Formato: | Artículo |
| Lenguaje: | Inglés |
| Publicado: |
Elsevier
2022
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12123/12906 https://www.sciencedirect.com/science/article/pii/S0167587722001738 https://doi.org/10.1016/j.prevetmed.2022.105740 |
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| author | Novoa, María Belén Aguirre, Nerina Valentini, Beatriz Susana Torioni, Susana Marta Signorini Porchiett, Marcelo Lisandro Primo, María Evangelina Elena, Sebastián Alejandro Vanzini, Victor Rene |
| author_browse | Aguirre, Nerina Elena, Sebastián Alejandro Novoa, María Belén Primo, María Evangelina Signorini Porchiett, Marcelo Lisandro Torioni, Susana Marta Valentini, Beatriz Susana Vanzini, Victor Rene |
| author_facet | Novoa, María Belén Aguirre, Nerina Valentini, Beatriz Susana Torioni, Susana Marta Signorini Porchiett, Marcelo Lisandro Primo, María Evangelina Elena, Sebastián Alejandro Vanzini, Victor Rene |
| author_sort | Novoa, María Belén |
| collection | INTA Digital |
| description | Brucellosis is an abortigenic and zoonotic disease. In cattle, it is mainly caused by Brucella abortus. The disease is endemic in low- and middle-income countries, being considered a neglected zoonotic disease. In these countries, it is of high importance to develop and validate sensitive, specific and low-cost diagnostic assays for brucellosis. The aim of the present study was the development of an indirect enzyme-linked immune assay (iELISA) to detect anti-B. abortus antibodies in milk samples. We purified the lipopolysaccharide antigen from B. abortus and produced an anti-bovine IgG monoclonal antibody to develop an iELISA (iELISAINTA). The iELISAINTA was validated using 1730 bulk milk samples and 1734 individual milk samples. The sampled dairy herds had at least 3 years of consistency at their positive or negative official brucellosis status. Individual milk samples were taken in parallel with serum samples from the cows. The status of the cows was defined by the result of the complement fixation test (CFT) performed with their serum sample. The reproducibility of the assay was evaluated in two laboratories. In addition, we evaluated the performance of the assay in the field, using 4385 bulk milk samples and 968 individual milk samples. The results of the iELISAINTA were compared with those obtained using the officially accepted brucellosis techniques: iELISA from Canada (iELISACFIA) in milk samples, and the buffered plate antigen (BPA) and the CFT in serum samples. At validation, the sensitivity (Se) of the iELISAINTA in bulk milk samples was 98.61 %, and the specificity (Sp) 98.79 % with a ≥ 10 % of positivity (PP) cutoff. In individual milk samples, the Se was 98.04 %, and the Sp 98.56 % with a ≥ 16 PP cutoff. The chance-corrected agreement kappa value (κ) between the results obtained in the different laboratories was κ = 0.87. In the field evaluation, in bulk milk samples the κ value between the iELISAINTA and the iELISACFIA was κ = 0.86. On individual milk samples, the κ values were: between the iELISAINTA and the iELISACFIA κ = 0.79, between the iELISAINTA and BPA was κ = 0.85, and between the iELISAINTA and CFT κ = 0.82. The developed iELISAINTA showed a very good performance and it could be used as a screening assay for anti-B. abortus antibodies detection in individual milk samples and for epidemiologic surveillance in bulk milk samples. |
| format | Artículo |
| id | INTA12906 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2022 |
| publishDateRange | 2022 |
| publishDateSort | 2022 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | INTA129062023-01-13T15:06:07Z Development, validation and field evaluation of an indirect ELISA for the detection of antibodies against Brucella abortus in bulk and individual milk samples in dairy cattle Novoa, María Belén Aguirre, Nerina Valentini, Beatriz Susana Torioni, Susana Marta Signorini Porchiett, Marcelo Lisandro Primo, María Evangelina Elena, Sebastián Alejandro Vanzini, Victor Rene Brucella abortus Brucelosis Anticuerpos Diagnóstico Ganado de Leche Brucellosis Antibodies Diagnosis Dairy Cattle Indirect ELISA ELISA Indirecto Brucellosis is an abortigenic and zoonotic disease. In cattle, it is mainly caused by Brucella abortus. The disease is endemic in low- and middle-income countries, being considered a neglected zoonotic disease. In these countries, it is of high importance to develop and validate sensitive, specific and low-cost diagnostic assays for brucellosis. The aim of the present study was the development of an indirect enzyme-linked immune assay (iELISA) to detect anti-B. abortus antibodies in milk samples. We purified the lipopolysaccharide antigen from B. abortus and produced an anti-bovine IgG monoclonal antibody to develop an iELISA (iELISAINTA). The iELISAINTA was validated using 1730 bulk milk samples and 1734 individual milk samples. The sampled dairy herds had at least 3 years of consistency at their positive or negative official brucellosis status. Individual milk samples were taken in parallel with serum samples from the cows. The status of the cows was defined by the result of the complement fixation test (CFT) performed with their serum sample. The reproducibility of the assay was evaluated in two laboratories. In addition, we evaluated the performance of the assay in the field, using 4385 bulk milk samples and 968 individual milk samples. The results of the iELISAINTA were compared with those obtained using the officially accepted brucellosis techniques: iELISA from Canada (iELISACFIA) in milk samples, and the buffered plate antigen (BPA) and the CFT in serum samples. At validation, the sensitivity (Se) of the iELISAINTA in bulk milk samples was 98.61 %, and the specificity (Sp) 98.79 % with a ≥ 10 % of positivity (PP) cutoff. In individual milk samples, the Se was 98.04 %, and the Sp 98.56 % with a ≥ 16 PP cutoff. The chance-corrected agreement kappa value (κ) between the results obtained in the different laboratories was κ = 0.87. In the field evaluation, in bulk milk samples the κ value between the iELISAINTA and the iELISACFIA was κ = 0.86. On individual milk samples, the κ values were: between the iELISAINTA and the iELISACFIA κ = 0.79, between the iELISAINTA and BPA was κ = 0.85, and between the iELISAINTA and CFT κ = 0.82. The developed iELISAINTA showed a very good performance and it could be used as a screening assay for anti-B. abortus antibodies detection in individual milk samples and for epidemiologic surveillance in bulk milk samples. EEA Rafaela Fil: Novoa, María Belen. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina Fil: Novoa, María Belen. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. 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. 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: Torioni, Susana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. 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: Signorini, Marcelo. 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: Primo, María Evangelina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina Fil: Elena, Sebastián. Servicio Nacional de Sanidad y Calidad Agroalimentaria. Dirección General de Laboratorio y Control Técnico. Laboratorio de Referencia de la OIE para Brucelosis; Argentina Fil: Vanzini, Victor Rene. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigación de la Cadena Láctea (IDICAL); Argentina Fil: Vanzini, Victor Rene. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Instituto de Investigación de la Cadena Láctea (IdICaL); Argentina 2022-09-16T12:54:00Z 2022-09-16T12:54:00Z 2022-11 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/12906 https://www.sciencedirect.com/science/article/pii/S0167587722001738 María Belén Novoa, Nerina Patricia Aguirre, Beatriz Valentini, Susana Torioni-de-Echaide, Marcelo Lisandro Signorini, María Evangelina Primo, Sebastián Elena, Victor René Vanzini, Development, validation and field evaluation of an indirect ELISA for the detection of antibodies against Brucella abortus in bulk and individual milk samples in dairy cattle, Preventive Veterinary Medicine, Volume 208, 2022, 105740, ISSN 0167-5877, https://doi.org/10.1016/j.prevetmed.2022.105740. 0167-5877 https://doi.org/10.1016/j.prevetmed.2022.105740 eng info:eu-repograntAgreement/INTA/2019-PE-E5-I109-001/2019-PE-E5-I109-001/AR./Convocatoria: Estudios para el control de enfermedades subtropicales y/o transmitidas por vectores (Tristeza Bovina, Garrapatas, Miasis, Tripanosomiasis, Lengua Azul y la info:eu-repograntAgreement/INTA/2019-RIST-E5-I111-001/2019-RIST-E5-I111-001/AR./Laboratorios de Diagnóstico Veterinario info:eu-repo/semantics/restrictedAccess application/pdf Elsevier Preventive Veterinary Medicine 208 : 105740 (November 2022) |
| spellingShingle | Brucella abortus Brucelosis Anticuerpos Diagnóstico Ganado de Leche Brucellosis Antibodies Diagnosis Dairy Cattle Indirect ELISA ELISA Indirecto Novoa, María Belén Aguirre, Nerina Valentini, Beatriz Susana Torioni, Susana Marta Signorini Porchiett, Marcelo Lisandro Primo, María Evangelina Elena, Sebastián Alejandro Vanzini, Victor Rene Development, validation and field evaluation of an indirect ELISA for the detection of antibodies against Brucella abortus in bulk and individual milk samples in dairy cattle |
| title | Development, validation and field evaluation of an indirect ELISA for the detection of antibodies against Brucella abortus in bulk and individual milk samples in dairy cattle |
| title_full | Development, validation and field evaluation of an indirect ELISA for the detection of antibodies against Brucella abortus in bulk and individual milk samples in dairy cattle |
| title_fullStr | Development, validation and field evaluation of an indirect ELISA for the detection of antibodies against Brucella abortus in bulk and individual milk samples in dairy cattle |
| title_full_unstemmed | Development, validation and field evaluation of an indirect ELISA for the detection of antibodies against Brucella abortus in bulk and individual milk samples in dairy cattle |
| title_short | Development, validation and field evaluation of an indirect ELISA for the detection of antibodies against Brucella abortus in bulk and individual milk samples in dairy cattle |
| title_sort | development validation and field evaluation of an indirect elisa for the detection of antibodies against brucella abortus in bulk and individual milk samples in dairy cattle |
| topic | Brucella abortus Brucelosis Anticuerpos Diagnóstico Ganado de Leche Brucellosis Antibodies Diagnosis Dairy Cattle Indirect ELISA ELISA Indirecto |
| url | http://hdl.handle.net/20.500.12123/12906 https://www.sciencedirect.com/science/article/pii/S0167587722001738 https://doi.org/10.1016/j.prevetmed.2022.105740 |
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