Background- Despite Brucellosis is eradicated from most of developed Countries, it still causes a public health impact and economic losses in animal husbandry in Mediterranean area. Nowadays, brucellosis is remerging as new challenge with outbreaks of both human and animal disease. Design of adequate strategies for preventing animal brucellosis and consequently human brucellosis requires understanding of microorganism and also of different strains. From diagnostic point of view, occurrence of false positive reactions in the serological tests available currently reduces their specificity. A “perfect” antigen has not been developed yet. 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. Proteome of Brucella spp is lack of consistency and so it is important set up protocols and methods to obtained proteins that are useful to develop rapid diagnostic test with immunoproteomics. Methods- 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. After 2D electrophoresis, image analysis was performed with Progenesis same spots and proteins were digested with trypsin and then analyzed with MALDI-TOF/MS (Bruker Daltonics). Results- 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. Conclusions- Proteomics is a useful approach to develop new diagnostic tools aimed to improve the outcome of Brucellosis eradication programs and to implement molecular epidemiology for a better understanding of spread and diffusion of brucellosis. Work supported by BrucMedNet- ARIMNet2 - Coordination of Agricultural Research in the Mediterranean grant agreement no. 618127.

New tools to improve serodiagnosis of Brucellosis / P. Roncada, A. Soggiu, C. Piras, V. Greco, T. Di Febo, I. Krasteva, A. Urbani, G. Garofolo, M. Tittarelli, L. Bonizzi. ((Intervento presentato al 16. convegno Human Proteome Organization World Congress tenutosi a Dublin nel 2017.

New tools to improve serodiagnosis of Brucellosis

A. Soggiu;C. Piras;L. Bonizzi
2017

Abstract

Background- Despite Brucellosis is eradicated from most of developed Countries, it still causes a public health impact and economic losses in animal husbandry in Mediterranean area. Nowadays, brucellosis is remerging as new challenge with outbreaks of both human and animal disease. Design of adequate strategies for preventing animal brucellosis and consequently human brucellosis requires understanding of microorganism and also of different strains. From diagnostic point of view, occurrence of false positive reactions in the serological tests available currently reduces their specificity. A “perfect” antigen has not been developed yet. 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. Proteome of Brucella spp is lack of consistency and so it is important set up protocols and methods to obtained proteins that are useful to develop rapid diagnostic test with immunoproteomics. Methods- 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. After 2D electrophoresis, image analysis was performed with Progenesis same spots and proteins were digested with trypsin and then analyzed with MALDI-TOF/MS (Bruker Daltonics). Results- 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. Conclusions- Proteomics is a useful approach to develop new diagnostic tools aimed to improve the outcome of Brucellosis eradication programs and to implement molecular epidemiology for a better understanding of spread and diffusion of brucellosis. Work supported by BrucMedNet- ARIMNet2 - Coordination of Agricultural Research in the Mediterranean grant agreement no. 618127.
giu-2017
Settore VET/05 - Malattie Infettive degli Animali Domestici
Settore VET/04 - Ispezione degli Alimenti di Origine Animale
Settore BIO/10 - Biochimica
New tools to improve serodiagnosis of Brucellosis / P. Roncada, A. Soggiu, C. Piras, V. Greco, T. Di Febo, I. Krasteva, A. Urbani, G. Garofolo, M. Tittarelli, L. Bonizzi. ((Intervento presentato al 16. convegno Human Proteome Organization World Congress tenutosi a Dublin nel 2017.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/556740
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