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

Bibliographic Details
Main Author: Baldani,Cristiane Divan
Publication Date: 2011
Other Authors: Hilario,Eduardo, Nakaghi,Andréa Cristina Higa, Bertolini,Maria Célia, Machado,Rosangela Zacarias
Format: Article
Language: eng
Source: Revista Brasileira de Parasitologia Veterinária (Online)
Download full: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1984-29612011000100011
Summary: 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 Escherichia 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 immuno assay in horses from São Paulo State, BrazilTheileria equidiagnosisEMA-1ELISAThe 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 Escherichia 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.Colégio Brasileiro de Parasitologia Veterinária2011-03-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1984-29612011000100011Revista Brasileira de Parasitologia Veterinária v.20 n.1 2011reponame:Revista Brasileira de Parasitologia Veterinária (Online)instname:Colégio Brasileiro de Parasitologia Veterinária (CBPV)instacron:CBPV10.1590/S1984-29612011000100011info:eu-repo/semantics/openAccessBaldani,Cristiane DivanHilario,EduardoNakaghi,Andréa Cristina HigaBertolini,Maria CéliaMachado,Rosangela Zacariaseng2011-05-05T00:00:00Zoai:scielo:S1984-29612011000100011Revistahttp://www.scielo.br/scielo.php?script=sci_serial&lng=pt&pid=1984-2961https://old.scielo.br/oai/scielo-oai.php||zacariascbpv@fcav.unesp.br1984-29610103-846Xopendoar:2011-05-05T00:00Revista Brasileira de Parasitologia Veterinária (Online) - Colégio Brasileiro de Parasitologia Veterinária (CBPV)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 immuno assay in horses from São Paulo State, Brazil
title Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immuno assay 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 immuno assay in horses from São Paulo State, Brazil
Baldani,Cristiane Divan
Theileria equi
diagnosis
EMA-1
ELISA
title_short Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immuno assay 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 immuno assay 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 immuno assay 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 immuno assay 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 immuno assay in horses from São Paulo State, Brazil
author Baldani,Cristiane Divan
author_facet Baldani,Cristiane Divan
Hilario,Eduardo
Nakaghi,Andréa Cristina Higa
Bertolini,Maria Célia
Machado,Rosangela Zacarias
author_role author
author2 Hilario,Eduardo
Nakaghi,Andréa Cristina Higa
Bertolini,Maria Célia
Machado,Rosangela Zacarias
author2_role author
author
author
author
dc.contributor.author.fl_str_mv Baldani,Cristiane Divan
Hilario,Eduardo
Nakaghi,Andréa Cristina Higa
Bertolini,Maria Célia
Machado,Rosangela Zacarias
dc.subject.por.fl_str_mv Theileria equi
diagnosis
EMA-1
ELISA
topic Theileria equi
diagnosis
EMA-1
ELISA
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 Escherichia 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 2011-03-01
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dc.publisher.none.fl_str_mv Colégio Brasileiro de Parasitologia Veterinária
publisher.none.fl_str_mv Colégio Brasileiro de Parasitologia Veterinária
dc.source.none.fl_str_mv Revista Brasileira de Parasitologia Veterinária v.20 n.1 2011
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