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(Reference retrieved automatically from SciELO through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

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Author(s):
Cristiane Divan Baldani [1] ; Eduardo Hilario [2] ; Andréa Cristina Higa Nakaghi [3] ; Maria Célia Bertolini [4] ; Rosangela Zacarias Machado [5]
Total Authors: 5
Affiliation:
[1] Universidade Federal Rural do Rio de Janeiro. Instituto de Veterinária. Departamento de Medicina e Cirurgia Veterinária
[2] University of California-Riverside. Department of Biochemistry
[3] Universidade Estadual Paulista. Faculdade de Ciências Agrárias e Veterinárias. Departamento de Patologia Veterinária
[4] Universidade Estadual Paulista. Departamento de Bioquímica e Tecnologia Química
[5] Universidade Estadual Paulista. Faculdade de Ciências Agrárias e Veterinárias. Departamento de Patologia Veterinária
Total Affiliations: 5
Document type: Journal article
Source: Revista Brasileira de Parasitologia Veterinária; v. 20, n. 1, p. 54-60, 2011-03-00.
Abstract

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. (AU)