Recombinant OMP28 antigen-based indirect ELISA for serodiagnosis of bovine brucellosis.
Brucellosis is a zoonosis of both public health and economic importance in many developing countries including India. Early detection and segregation of the infected animals are important in order to control the disease. Serodiagnostic tests for brucellosis is mainly based on detection of antibodies developed against lipopolysaccharide (LPS) component of cell. In this study we evaluated a protein antigen, 28 kDa outer membrane protein (OMP28), of Brucella melitensis as an alternative to LPS. Recombinant OMP28 was produced in Escherichia coli system. The efficacy of purified OMP28 was studied in an indirect enzyme-linked immunosorbent assay (ELISA) for diagnosis of brucellosis in field sera collected from different regions of country. Using known negative and known positive serum samples it was found that OMP28 is immunoreactive to Brucella infected cattle, sheep, goat and dog sera. Three hundred and eighty two cattle sera were screened by OMP28 antigen-based ELISA and the results were compared to rose Bengal plate agglutination Test (RBPT). Recombinant OMP28 antigen-based ELISA has shown sensitivity of 88.7%, specificity of 93.8% and accuracy of 92.9%. It was concluded that recombinant B. melitensis OMP28 could be used as a protein antigen for diagnosis of brucellosis in domestic animals.
Chaudhuri
Pallab
P
Genetic Engineering of Bacteria Lab., Division of Bacteriology, Indian Veterinary Research Institute, Izatnagar, India. pallab.chaudhuri@gmail.com
Prasad
Rajeev
R
Kumar
Vinoth
V
Gangaplara
Arunakumar
A
D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D017347:Brucella melitensis / Q000276:immunology*; D002007:Brucellosis, Bovine / Q000097:blood / Q000175:diagnosis* / Q000276:immunology; D002417:Cattle; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D006041:Goats; D006801:Humans; D007106:Immune Sera / Q000097:blood / Q000276:immunology; D015151:Immunoblotting; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D012680:Sensitivity and Specificity; D012698:Serologic Tests
D000942:Antigens, Bacterial; C106040:BP26 protein, Brucella; D007106:Immune Sera; D008565:Membrane Proteins; D011994:Recombinant Proteins