An animal model of tick-borne encephalitis virus (TBEV) infection was established to provide a suitable tool for anti-TBEV drug screening. BALB/c mice were inoculated with TBEV by subcutaneous injection, and their symptoms, body weight and survival rate were observed. TBEV antigen distribution, virus titer, pathological changes and the expression levels of tumor necrosis factor α (TNF-α) and interferon β (IFN-β) mRNA were investigated in brain, spleen and kidney of the mice. The results showed that the infected mice had symptoms of arched back and hind limb paralysis. The body weight and survival rate of infected mice were significantly decreased as compared with the uninfected mice. TBEV antigen could be detected in the brain, spleen and kidney of infected mice. Virus titers in the brain were higher than those in the spleen and kidney of infected mice, along with the pathological changes in the brain. The expressions of TNF-α and IFN-β mRNA in the brain, spleen and kidney of mice were associated with the duration of TBEV infection. BALB/c mice model of TBEV infection has been established by subcutaneous injection of virus.