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Development of an indirect ELISA based on recombinant capsid protein to detect antibodies to bovine leukemia virus

Serological testing and culling infected animals are key management practices aiming eradication of bovine leukemia virus infection. Here, we report the development of an indirect ELISA based on BLV recombinant capsid protein (BLVp24r) to detect anti-BLV antibodies in cattle serum. The BLVp24r was e...

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Autores principales: Andreolla, Ana Paula, Erpen, Luana Marina Scheer, Frandoloso, Rafael, Kreutz, Luiz Carlos
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6328717/
https://www.ncbi.nlm.nih.gov/pubmed/29866609
http://dx.doi.org/10.1016/j.bjm.2018.05.001
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author Andreolla, Ana Paula
Erpen, Luana Marina Scheer
Frandoloso, Rafael
Kreutz, Luiz Carlos
author_facet Andreolla, Ana Paula
Erpen, Luana Marina Scheer
Frandoloso, Rafael
Kreutz, Luiz Carlos
author_sort Andreolla, Ana Paula
collection PubMed
description Serological testing and culling infected animals are key management practices aiming eradication of bovine leukemia virus infection. Here, we report the development of an indirect ELISA based on BLV recombinant capsid protein (BLVp24r) to detect anti-BLV antibodies in cattle serum. The BLVp24r was expressed in Escherichia coli and purified by affinity chromatography, and then used to set up the ELISA parameters. The Polysorp(®) plate coated with 50 ng of antigen/well and bovine serum diluted 1:100 gave the best results during standardization. Using sera from infected and non-infected cattle we set up the cutoff point at 0.320 (OD(450 nm)) with a sensitivity of 98.5% and specificity of 100.0%. Then, we tested 1.187 serum samples from dairy (736 samples) and beef cattle (451 samples) with unknown status to BLV. We found that 31.1% (229/736) and 9.5% (43/451) of samples amongst dairy and beef cattle, respectively, had IgGs to BLV. The rate of agreement with a commercial competitive ELISA was 84.3% with a κ value of 0.68. Thus, our BLVp24r iELISA is suitable to detect BLV infected animals and should be a useful tool to control BLV infection in cattle.
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spelling pubmed-63287172019-01-22 Development of an indirect ELISA based on recombinant capsid protein to detect antibodies to bovine leukemia virus Andreolla, Ana Paula Erpen, Luana Marina Scheer Frandoloso, Rafael Kreutz, Luiz Carlos Braz J Microbiol Research Paper Serological testing and culling infected animals are key management practices aiming eradication of bovine leukemia virus infection. Here, we report the development of an indirect ELISA based on BLV recombinant capsid protein (BLVp24r) to detect anti-BLV antibodies in cattle serum. The BLVp24r was expressed in Escherichia coli and purified by affinity chromatography, and then used to set up the ELISA parameters. The Polysorp(®) plate coated with 50 ng of antigen/well and bovine serum diluted 1:100 gave the best results during standardization. Using sera from infected and non-infected cattle we set up the cutoff point at 0.320 (OD(450 nm)) with a sensitivity of 98.5% and specificity of 100.0%. Then, we tested 1.187 serum samples from dairy (736 samples) and beef cattle (451 samples) with unknown status to BLV. We found that 31.1% (229/736) and 9.5% (43/451) of samples amongst dairy and beef cattle, respectively, had IgGs to BLV. The rate of agreement with a commercial competitive ELISA was 84.3% with a κ value of 0.68. Thus, our BLVp24r iELISA is suitable to detect BLV infected animals and should be a useful tool to control BLV infection in cattle. Elsevier 2018-05-22 /pmc/articles/PMC6328717/ /pubmed/29866609 http://dx.doi.org/10.1016/j.bjm.2018.05.001 Text en © 2018 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Research Paper
Andreolla, Ana Paula
Erpen, Luana Marina Scheer
Frandoloso, Rafael
Kreutz, Luiz Carlos
Development of an indirect ELISA based on recombinant capsid protein to detect antibodies to bovine leukemia virus
title Development of an indirect ELISA based on recombinant capsid protein to detect antibodies to bovine leukemia virus
title_full Development of an indirect ELISA based on recombinant capsid protein to detect antibodies to bovine leukemia virus
title_fullStr Development of an indirect ELISA based on recombinant capsid protein to detect antibodies to bovine leukemia virus
title_full_unstemmed Development of an indirect ELISA based on recombinant capsid protein to detect antibodies to bovine leukemia virus
title_short Development of an indirect ELISA based on recombinant capsid protein to detect antibodies to bovine leukemia virus
title_sort development of an indirect elisa based on recombinant capsid protein to detect antibodies to bovine leukemia virus
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6328717/
https://www.ncbi.nlm.nih.gov/pubmed/29866609
http://dx.doi.org/10.1016/j.bjm.2018.05.001
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