Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immunoassay in horses from São Paulo State, Brazil

Detalhes bibliográficos
Autor(a) principal: Baldani, Cristiane Divan
Data de Publicação: 2011
Outros Autores: Hilario, Eduardo, Nakaghi, Andréa Cristina Higa [UNESP], Bertolini, Maria Célia [UNESP], Machado, Rosangela Zacarias [UNESP]
Tipo de documento: Artigo
Idioma: por
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1590/s1984-29612011000100011
http://hdl.handle.net/11449/226311
Resumo: The erythrocytic-stage surface protein, Equi Merozoite Antigen 1 (EMA-1), is a major candidate for the development of a diagnostic antigen for equine piroplasmosis. In order to establish an effective diagnostic method for practical use, the gene encoding the entire EMA-1 of Theileria equi Jaboticabal strain was cloned and expressed in scherichia coli as a histidine-tagged protein (His6-EMA1). The expressed EMA-1 reacted with specific antibodies in Western blot and had an apparent molecular mass of 34 kDa which was largely consistent with its theoretical value. The nucleotide sequence of the EMA-1 gene of Jaboticabal strain was comparatively analyzed with other published sequences. The results indicated a high degree of homology with EMA-1 genes of all other strains isolated from various countries. The recombinant purified His6-EMA1 protein was tested in an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies anti-T. equi in horses. The ELISA clearly differentiated T. equi-infected from Babesia caballi-infected horse sera or normal horse sera. Field serum samples collected from horses in the State of São Paulo, Southeastern Brazil, were examined for the diagnosis of T. equi infection by ELISA. Of 170 samples analyzed, 95.88% (163/170) were positive for T. equi infection. These results suggest that the His6-EMA1 protein expressed in E. coli could be a reliable immunodiagnostic antigen for ELISA test and that T. equi infection is a serious concern in the State of São Paulo, Brazil.
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spelling Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immunoassay in horses from São Paulo State, BrazilProdução da proteína recombinante EMA-1 e sua aplicação para o diagnóstico baseado no imunoensaio enzimático de Theileria equi em equinos do Estado de São Paulo, BrasilDiagnosisELISAEMA-1Theileria equiThe erythrocytic-stage surface protein, Equi Merozoite Antigen 1 (EMA-1), is a major candidate for the development of a diagnostic antigen for equine piroplasmosis. In order to establish an effective diagnostic method for practical use, the gene encoding the entire EMA-1 of Theileria equi Jaboticabal strain was cloned and expressed in scherichia coli as a histidine-tagged protein (His6-EMA1). The expressed EMA-1 reacted with specific antibodies in Western blot and had an apparent molecular mass of 34 kDa which was largely consistent with its theoretical value. The nucleotide sequence of the EMA-1 gene of Jaboticabal strain was comparatively analyzed with other published sequences. The results indicated a high degree of homology with EMA-1 genes of all other strains isolated from various countries. The recombinant purified His6-EMA1 protein was tested in an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies anti-T. equi in horses. The ELISA clearly differentiated T. equi-infected from Babesia caballi-infected horse sera or normal horse sera. Field serum samples collected from horses in the State of São Paulo, Southeastern Brazil, were examined for the diagnosis of T. equi infection by ELISA. Of 170 samples analyzed, 95.88% (163/170) were positive for T. equi infection. These results suggest that the His6-EMA1 protein expressed in E. coli could be a reliable immunodiagnostic antigen for ELISA test and that T. equi infection is a serious concern in the State of São Paulo, Brazil.Departamento de Medicina e Cirurgia Veterinária Instituto de Veterinária Universidade Federal Rural do Rio de Janeiro - UFRRJDepartment of Biochemistry University of California-Riverside, Riverside, CADepartamento de Patologia Veterinária Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista - UNESPDepartamento de Bioquímica e Tecnologia Química Universidade Estadual Paulista - UNESPDepartamento de Patologia Veterinária Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista - UNESPDepartamento de Bioquímica e Tecnologia Química Universidade Estadual Paulista - UNESPUniversidade Federal Rural do Rio de Janeiro - UFRRJUniversity of California-RiversideUniversidade Estadual Paulista (UNESP)Baldani, Cristiane DivanHilario, EduardoNakaghi, Andréa Cristina Higa [UNESP]Bertolini, Maria Célia [UNESP]Machado, Rosangela Zacarias [UNESP]2022-04-28T22:37:16Z2022-04-28T22:37:16Z2011-01-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article54-60http://dx.doi.org/10.1590/s1984-29612011000100011Revista Brasileira de Parasitologia Veterinaria, v. 20, n. 1, p. 54-60, 2011.0103-846Xhttp://hdl.handle.net/11449/22631110.1590/s1984-296120110001000112-s2.0-79955144602Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPporRevista Brasileira de Parasitologia Veterinariainfo:eu-repo/semantics/openAccess2024-06-07T13:01:16Zoai:repositorio.unesp.br:11449/226311Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T13:35:06.193992Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immunoassay in horses from São Paulo State, Brazil
Produção da proteína recombinante EMA-1 e sua aplicação para o diagnóstico baseado no imunoensaio enzimático de Theileria equi em equinos do Estado de São Paulo, Brasil
title Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immunoassay in horses from São Paulo State, Brazil
spellingShingle Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immunoassay in horses from São Paulo State, Brazil
Baldani, Cristiane Divan
Diagnosis
ELISA
EMA-1
Theileria equi
title_short Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immunoassay in horses from São Paulo State, Brazil
title_full Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immunoassay in horses from São Paulo State, Brazil
title_fullStr Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immunoassay in horses from São Paulo State, Brazil
title_full_unstemmed Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immunoassay in horses from São Paulo State, Brazil
title_sort Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immunoassay in horses from São Paulo State, Brazil
author Baldani, Cristiane Divan
author_facet Baldani, Cristiane Divan
Hilario, Eduardo
Nakaghi, Andréa Cristina Higa [UNESP]
Bertolini, Maria Célia [UNESP]
Machado, Rosangela Zacarias [UNESP]
author_role author
author2 Hilario, Eduardo
Nakaghi, Andréa Cristina Higa [UNESP]
Bertolini, Maria Célia [UNESP]
Machado, Rosangela Zacarias [UNESP]
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Universidade Federal Rural do Rio de Janeiro - UFRRJ
University of California-Riverside
Universidade Estadual Paulista (UNESP)
dc.contributor.author.fl_str_mv Baldani, Cristiane Divan
Hilario, Eduardo
Nakaghi, Andréa Cristina Higa [UNESP]
Bertolini, Maria Célia [UNESP]
Machado, Rosangela Zacarias [UNESP]
dc.subject.por.fl_str_mv Diagnosis
ELISA
EMA-1
Theileria equi
topic Diagnosis
ELISA
EMA-1
Theileria equi
description The erythrocytic-stage surface protein, Equi Merozoite Antigen 1 (EMA-1), is a major candidate for the development of a diagnostic antigen for equine piroplasmosis. In order to establish an effective diagnostic method for practical use, the gene encoding the entire EMA-1 of Theileria equi Jaboticabal strain was cloned and expressed in scherichia coli as a histidine-tagged protein (His6-EMA1). The expressed EMA-1 reacted with specific antibodies in Western blot and had an apparent molecular mass of 34 kDa which was largely consistent with its theoretical value. The nucleotide sequence of the EMA-1 gene of Jaboticabal strain was comparatively analyzed with other published sequences. The results indicated a high degree of homology with EMA-1 genes of all other strains isolated from various countries. The recombinant purified His6-EMA1 protein was tested in an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies anti-T. equi in horses. The ELISA clearly differentiated T. equi-infected from Babesia caballi-infected horse sera or normal horse sera. Field serum samples collected from horses in the State of São Paulo, Southeastern Brazil, were examined for the diagnosis of T. equi infection by ELISA. Of 170 samples analyzed, 95.88% (163/170) were positive for T. equi infection. These results suggest that the His6-EMA1 protein expressed in E. coli could be a reliable immunodiagnostic antigen for ELISA test and that T. equi infection is a serious concern in the State of São Paulo, Brazil.
publishDate 2011
dc.date.none.fl_str_mv 2011-01-01
2022-04-28T22:37:16Z
2022-04-28T22:37:16Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1590/s1984-29612011000100011
Revista Brasileira de Parasitologia Veterinaria, v. 20, n. 1, p. 54-60, 2011.
0103-846X
http://hdl.handle.net/11449/226311
10.1590/s1984-29612011000100011
2-s2.0-79955144602
url http://dx.doi.org/10.1590/s1984-29612011000100011
http://hdl.handle.net/11449/226311
identifier_str_mv Revista Brasileira de Parasitologia Veterinaria, v. 20, n. 1, p. 54-60, 2011.
0103-846X
10.1590/s1984-29612011000100011
2-s2.0-79955144602
dc.language.iso.fl_str_mv por
language por
dc.relation.none.fl_str_mv Revista Brasileira de Parasitologia Veterinaria
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 54-60
dc.source.none.fl_str_mv Scopus
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
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