Lala Lajpat Rai University of Veterinary & Animal Sciences, Hisar

Browse

Now showing 1 - 1 of 1

Restricted
Development of Recombinant antigen based ELISA for Serodiagnosis of Theileria equi in equines
(LUVAS, 2012) Sandeep; Rakha, N.K.
The erythrocytic-stage surface protein, Equi Merozoite Antigen 1 (EMA-1), is a major candidate for the development of a immunodiagnostic antigen forTheileria equiinfection . In order to establish an effective diagnostic method for laboratory use, 547bp gene segment of EMA-1 of Theileria equi from Indian isolate was expressed in Escherichia coli as a Glutathione-S-transferase-tagged protein (GSTEMA-1) and the fusion protein was purified by Glutathione sepharose beads. The expressed fusion protein reacted with specific antibodies in Western blot and had an apparent molecular mass of 43 kDa which was largely consistent with its theoretical value. The recombinant purified GST-EMA-1 protein was used in enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against T. equi in horses. The ELISA clearly differentiated T. equi-infected from normal horse sera. Field serum samples collected from horses in Haryana and Uttarakhand, were examined for the determination of T. equi antibodies in ELISA. Of 240 samples analyzed, 45.41% (106/240) were positive for T. equi infection. These results suggest that the GST-EMA-1 protein expressed in E. coli could be a reliable immunodiagnostic antigen for ELISA and that T. equi infection is endemic in the areas from which samples were collected.