Objective To establish a HFMD BALB/ c sulking mouse model infected by coxsackievirus A16, and evaluate immunological and pathological characteristics of this model. Methods In this study, 3-day-old BALB/ c suckling mice were infected with 100 μL coxsackievirus A16 via intraperitoneal injection. The survival rate, clinical score and the 50% lethal dose (LD₅₀ ) were monitored daily 14 days after infection. Challenge with the coxsackievirus A16 with a dose of 3LD₅₀ , 3-day-old BALB/ c suckling mice were monitored daily the survival rate, clinical score and weight within 14 d. 6 d after infection, the concentration of MCP-1, MIP-1 and G-CSF in serum was determined by cytometric bead array (CBA) method ; viral loads of the hind limb muscle, heart, brain, intestine were determined by the real-time RT-PCR; histological change of muscle and brain were determined. Results After different concentrations of virus infection, 3-day- old BALB/ c suckling mice appeared inactivity, hind limb weakness, paralysis and even death, and the virulence of coxsackievirus A16 for 3-day-old BALB/ c suckling mice was 56 LD₅₀ / mL. Within 14 days after infection, compared with the control group, there were significant differences in the survival days, mortality, clinical score, and body weight of suckling mice in the model group (P< 0. 01).6 d post infection, the concentrations of MCP-1 and G-CSF in serum are significantly increased (P< 0. 01) compared with the control group. The viral loads of hindlimb muscle, heart, brain and intestine was significantly higher than that of the control group (P< 0. 01), and the viral loads of hindlimb muscle was the highest. Pathological change showed that there were large area of atrophy and inflammation in the hind limbs muscle and atrophy in the nerve cells of the brain. Conclusions A CoxA16 infected 3-day-old BALB/ c suckling mouse model was successfully established, which can serve for medicine evaluation and mechanism.