Abstract—Human vaccines for brucellosis are still not licensed and in some cases, animal vaccines are used for humans. We herein conducted a study to identify and characterize a suitable T cell epitope, which might be efficient against Brucella spp. We have assessed all the antigenic outer membrane proteins of Brucella spp recorded to date in Protegen (http://www.violinet.org/protegen/ index.php) database and found that outer membrane protein (Omp31) of Brucella melitensis M28 was superior among all others. Analysis of Omp31 of Brucella melitensis M28 with NetMHCII 2.2, SYFPEITHI, ProPred and NetCTL 1.2 servers for identification of T cell epitope predicted an epitope 125-VRARLGYTATERLMV-139. These four servers also predicted that the epitope could bind with at least 6 major histocompatibility complex class-II (MHC II) and 5 MHC I molecules. The epitope shared ~86.93% identity with Omp31 of all human antigenic Brucella spp. Analysis with different bioinformatics tools revealed that this epitope was highly stable and capable to induce T cell-mediated immunity. Finally, molecular docking simulation showed that it could bind with the binding groove of MHC II and MHC I molecules with numerous significant hydrogen bonds. Results reported herein indicate that 125-VRARLGYTATERLMV-139 might be the target for epitope-based vaccine in humans against brucellosis.