Abstract
Background: The diagnosis of brucellosis by serological tests is based on antigen suspensions derived from smooth lipopolysaccharide extracts, which can give false positive results linked to cross-reactivity with other Gram-negative microorganisms, especially Yersinia enterocolitica O:9 and Escherichia coli O157:H7. Objective: The objective of the present study was the characterization by proteomic analysis of specific immunogenic proteins not associated with smooth lipopolysaccharide to improve the diagnostic tests used in the ovine brucellosis eradication programs. Methods: The serum from a sheep positive to Brucella melitensis was treated to eliminate all antibodies against such lipopolysaccharide and highlight the reaction towards the immunoreactive proteins in Western Blotting. Results: The immunoreactive bands were identified by nLC-MS/MS and through bioinformatic tools, it was possible to select 12 potential candidates as protein antigens specific for Brucella melitensis. Conclusion: The detection of new antigens not subjected to cross-reactivity with other Gram-negative microorganisms can offer an additional tool for the serological diagnosis of such disease.
Highlights
The brucellosis can affect humans by the contact with ill animals and their parts, such as carcasses or abortion derivatives, and by consumption of contaminated raw milk and unpasteurized dairy products [1]
The immunoreactive bands were identified by nLC-MS/MS and through bioinformatic tools, it was possible to select 12 potential candidates as protein antigens specific for Brucella melitensis
Brucella melitensis (BM) immunogenic proteins were investigated to detect epitopes able to distinguish true BM positive animals from those infected with Y. enterocolitica O:9 and E. coli O157:H7, the most important Brucella cross-reactive bacteria
Summary
The brucellosis can affect humans by the contact with ill animals and their parts, such as carcasses or abortion derivatives, and by consumption of contaminated raw milk and unpasteurized dairy products [1]. The first step was the localization of proteins to discard the cytosolic ones, because the proteins of the envelope are those that are into contact with the host immune system and stimulate the production of antibodies Such envelope proteins were studied for their immunogenic potential with the aim to discard the homologue proteins with the major cross-reactive Gramnegative bacteria. The diagnosis of brucellosis by serological tests is based on antigen suspensions derived from smooth lipopolysaccharide extracts, which can give false positive results linked to cross-reactivity with other Gram-negative microorganisms, especially Yersinia enterocolitica O:9 and Escherichia coli O157:H7
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