Proteomics to improve serodiagnosis of Brucellosis

Brucellosis still causes a public health impact and economic losses in animal husbandry in Mediterranean area also if it is eradicated in most of developed Countries. The design of adequate strategies for preventing animal brucellosis and consequently human disease requires in-depth knowledge of different strains that are circulating. Moreover, the occurrence of false positive reactions in the serological tests available currently reduces their specificity [1]. Furthermore, the mass vaccination policy using Rev.1 vaccine, in many endemic areas of the Mediterranean with the aim of controlling/reducing disease prevalence in small ruminants, interfere with serological diagnosis of the disease. Since the 'perfect antigen' has not been developed yet, it is useful to make comparative proteomics characterization of of Brucella spp to set to obtained proteins that could permit to develop rapid diagnostic test. We analyze three strains of Brucella melitensis coming from library of National Reference Laboratory of Brucellosis (IZSAM- Teramo ITALY) in three technical replicates for each strain used a combined methods of bead beating and solubilization [2]. After 2D electrophoresis, image analysis was performed with Progenesis same spots and proteins were analyzed with MALDI-TOF/MS. Combined methods of extraction physical plus chemical were able to resolve huge numbers of proteins that can be used to set up immune-proteomics analysis to improve and complete serodiagnosis of brucellosis Work supported by BrucMedNet- ARIMNet2 - Coordination of Agricultural Research in the Mediterranean-Grant agreement no. 618127. * Corresponding author: Paola Roncada, PhD. Istituto Sperimentale Italiano Lazzaro Spallanzani, Milano Phone.: +39 0250318138 E-mail address: paola.roncada@istitutospallanzani.it References: [1] Di Febo T, Luciani M, Portanti O, Bonfini B, Lelli R, Tittarelli M Development and evaluation of diagnostic tests for the serological diagnosis of brucellosis in swine. Vet Ital. 2012 Apr-Jun;48(2):133-56. [2] Piras C, Soggiu A, Bonizzi L, Greco V, Ricchi M, Arrigoni N, Bassols A, Urbani A, Roncada P.Identification of immunoreactive proteins