Development of an indirect ELISA based on recombinant capsid protein to detect antibodies to bovine leukemia virus
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Outros Autores: | , , |
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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Brazilian Journal of Microbiology |
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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 |
_version_ |
1752122209828077568 |