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Page No. : 117-121

ABSTRACT

Brucellosis in man and animals remains an important zoonosis all over the world. An ideal diagnostic test and protective vaccine are essential to control and eradicate the malady which requires identification of a suitable antigen. A soluble antigen from Brucella melitensis 16M (BMSA) was prepared and immunochemically characterized. The BmSA contained 2.75 mg/ml of protein and 1.52 mg/ml of carbohydrate. The SDS-PAGE on 12.5% acrylamide gel of BmSA produced peptide bands in the range of 95 to 13 kDa molecular weight. The immunodominant peptides in the BmSA were identified by Western blot analysis using anti-B. melitensis 16M rabbit serum, anti-B. melitensis Rev. 1 vaccinated sheep serum, anti-B. abortus bovine serum and clinical serum from human. The serum samples produced different bands. The anti-B. melitensis 16M rabbit serum produced bands of 39, 20 and 18 kDa, whereas the Rev. 1 vaccinated sheep serum produced immunodominant bands of 18-13 kDa. The anti-B. abortus bovine serum produced bands of 39-29 and 13 kDa while very faint bands of 43 and 13 kDa were observed with human clinical serum. The low molecular weight peptides of BmSA were more immunodominant.

Keywords: Brucellosis, Brucella melitensis, sheep, SDS-PAGE, Western blotting.