Background: Brucellosis remains a significant public health issue that affects the agricultural economy in Mediterranean basin. The disease is widespread zoonosis that infects humans mostly through the consumption of contaminated raw milk and dairy products or by contact with infected animals. Although several control programs have been implemented in the Mediterranean basin, the disease is still prevalent in small ruminants and further actions are required to improve the surveillance. Animal vaccination is an efficacious countermeasure but the current diagnostic tests do not allow the differentiation of infected from vaccinated animals (DIVA), limiting the intervention strategies to fight the disease. In this study, we performed an immunoproteomic characterization of Brucella melitensis vaccine strain Rev. 1 and the wild type strain 16M in order to identify candidate antigens specific for infection or vaccination. Methods: The B. melitensis vaccine strain Rev.1 and the wild-type strain16M have been subjected to optimized 2D western blot against immune sera from naturally infected animals using a combined fluorescence-chemilumiscence detection. Western blot (WB) image analysis has been performed with Image Studio 5 (LiCor) and specific immunoreactive proteins were identified by MALDI-TOF/TOF with LIFT (Bruker Daltonics). Results: Three immunoreactive proteins that specifically recognize 16M strain have been detected and identified by the optimized 2D-WB coupled to MALDI-MS/MS identification Conclusions: optimized High resolution 2DE and western blotting identified a panel of candidate antigens that could discriminate Infected from Vaccinated Animals, supporting the development of DIVA tests for an improved disease management. Brucmednet is a project funded by ARIMNet2, an ERA-NET supported and funded by the European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement no. 618127
Identification of New candidate antigens to support diva strategies in Brucellosis.
Piras C;Britti D;
2018-01-01
Abstract
Background: Brucellosis remains a significant public health issue that affects the agricultural economy in Mediterranean basin. The disease is widespread zoonosis that infects humans mostly through the consumption of contaminated raw milk and dairy products or by contact with infected animals. Although several control programs have been implemented in the Mediterranean basin, the disease is still prevalent in small ruminants and further actions are required to improve the surveillance. Animal vaccination is an efficacious countermeasure but the current diagnostic tests do not allow the differentiation of infected from vaccinated animals (DIVA), limiting the intervention strategies to fight the disease. In this study, we performed an immunoproteomic characterization of Brucella melitensis vaccine strain Rev. 1 and the wild type strain 16M in order to identify candidate antigens specific for infection or vaccination. Methods: The B. melitensis vaccine strain Rev.1 and the wild-type strain16M have been subjected to optimized 2D western blot against immune sera from naturally infected animals using a combined fluorescence-chemilumiscence detection. Western blot (WB) image analysis has been performed with Image Studio 5 (LiCor) and specific immunoreactive proteins were identified by MALDI-TOF/TOF with LIFT (Bruker Daltonics). Results: Three immunoreactive proteins that specifically recognize 16M strain have been detected and identified by the optimized 2D-WB coupled to MALDI-MS/MS identification Conclusions: optimized High resolution 2DE and western blotting identified a panel of candidate antigens that could discriminate Infected from Vaccinated Animals, supporting the development of DIVA tests for an improved disease management. Brucmednet is a project funded by ARIMNet2, an ERA-NET supported and funded by the European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement no. 618127I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
