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

Detalhes bibliográficos
Autor(a) principal: Andreolla,Ana Paula
Data de Publicação: 2018
Outros Autores: Erpen,Luana Marina Scheer, Frandoloso,Rafael, Kreutz,Luiz Carlos
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Brazilian Journal of Microbiology
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822018000500068
Resumo: Abstract 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 (OD450 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 Development of an indirect ELISA based on recombinant capsid protein to detect antibodies to bovine leukemia virusBovine leukemia virusRetrovirusDiagnosisELISAAbstract 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 (OD450 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.Sociedade Brasileira de Microbiologia2018-01-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822018000500068Brazilian Journal of Microbiology v.49 suppl.1 2018reponame:Brazilian Journal of Microbiologyinstname:Sociedade Brasileira de Microbiologia (SBM)instacron:SBM10.1016/j.bjm.2018.05.001info:eu-repo/semantics/openAccessAndreolla,Ana PaulaErpen,Luana Marina ScheerFrandoloso,RafaelKreutz,Luiz Carloseng2018-11-29T00:00:00Zoai:scielo:S1517-83822018000500068Revistahttps://www.scielo.br/j/bjm/ONGhttps://old.scielo.br/oai/scielo-oai.phpbjm@sbmicrobiologia.org.br||mbmartin@usp.br1678-44051517-8382opendoar:2018-11-29T00:00Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM)false
dc.title.none.fl_str_mv 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
spellingShingle Development of an indirect ELISA based on recombinant capsid protein to detect antibodies to bovine leukemia virus
Andreolla,Ana Paula
Bovine leukemia virus
Retrovirus
Diagnosis
ELISA
title_short 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_sort Development of an indirect ELISA based on recombinant capsid protein to detect antibodies to bovine leukemia virus
author Andreolla,Ana Paula
author_facet Andreolla,Ana Paula
Erpen,Luana Marina Scheer
Frandoloso,Rafael
Kreutz,Luiz Carlos
author_role author
author2 Erpen,Luana Marina Scheer
Frandoloso,Rafael
Kreutz,Luiz Carlos
author2_role author
author
author
dc.contributor.author.fl_str_mv Andreolla,Ana Paula
Erpen,Luana Marina Scheer
Frandoloso,Rafael
Kreutz,Luiz Carlos
dc.subject.por.fl_str_mv Bovine leukemia virus
Retrovirus
Diagnosis
ELISA
topic Bovine leukemia virus
Retrovirus
Diagnosis
ELISA
description Abstract 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 (OD450 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.
publishDate 2018
dc.date.none.fl_str_mv 2018-01-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822018000500068
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822018000500068
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1016/j.bjm.2018.05.001
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv text/html
dc.publisher.none.fl_str_mv Sociedade Brasileira de Microbiologia
publisher.none.fl_str_mv Sociedade Brasileira de Microbiologia
dc.source.none.fl_str_mv Brazilian Journal of Microbiology v.49 suppl.1 2018
reponame:Brazilian Journal of Microbiology
instname:Sociedade Brasileira de Microbiologia (SBM)
instacron:SBM
instname_str Sociedade Brasileira de Microbiologia (SBM)
instacron_str SBM
institution SBM
reponame_str Brazilian Journal of Microbiology
collection Brazilian Journal of Microbiology
repository.name.fl_str_mv Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM)
repository.mail.fl_str_mv bjm@sbmicrobiologia.org.br||mbmartin@usp.br
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