Use of Some Mycobacterial Peptides and Recombinant Proteins for Diagnosis of Bovine Tuberculosis

Abstract

With the procedure described in this work, it is possible to clone, express, purify successfully M. bovis ESAT-6, MPB-83 and MPB-64, in a recombinant form in E. coli cells, and use in vivo by measuring cell mediated immunity in guinea pigs using skin sensitivity test. SDS-PAGE was applied for characterization of recombinant proteins of M. bovis. rESAT-6 revealed protein at Mw of 14 kDa, rMPB-83 revealed proteins at Mw of 26 kDa, & rMPB-64 revealed proteins at Mw of 24 kDa. Western blotting analysis was performed, the results showed clear visible immune-reactive bands in rESAT-6 at Mw of 14 kDa, in rMPB-83 at 26 kDa, and in & rMPB-64 at 24 kDa. The obtained results of skin sensitivity test on M.bovis experimentally infected guinea pigs revealed that, each of M. bovis rESAT-6, rMPB-83 and rMPB-64 antigens could be able to synthetized and differentiate infected guinea pig groups from non-infected.