Identification of Immunogenic Candidate for New Serological Tests for Brucella melitensis by a Proteomic Approach

Valentina Paci1, Pierina Visciano2, Ivanka Krasteva1, Tiziana Di Febo1, Fabrizia Perletta1, Chiara Di Pancrazio1, Federica D’Onofrio2, Maria Schirone2, *, Manuela Tittarelli1, Mirella Luciani1
1 Istituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise “G. Caporale”, Via Campo Boario, 64100 Teramo, Italy
2 Faculty of Bioscience and Technology for Food, Agriculture and Environment, University of Teramo, Via R. Balzarini 1, 64100 Teramo, Italy

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© 2021 Paci et al.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the Faculty of Bioscience and Technology for Food, Agriculture and Environment - University of Teramo, Via R. Balzarini 1, 64100 Teramo, Italy; Tel: +390861 266911; E-mail:



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.


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.


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.


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.


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.

Keywords: Brucellosis, Cross-reactivity, Western blotting, Mass spectrometry, Bioinformatics, Serological tests.