Objective：To identify the HLAA0201 restricted CTL epitopes derived from hepatitis B virus X protein predicted by computer program and general principles in vitro. Methods: HBx gene sequences of Hepatitis B virus genotypes B/C and serotypes adw/adr,with the highest frequencies in Chinese,were computed and analyzed by screening service offered by Internet combined with peptide supermotif,extended motif and quantitative motif prediction. Four most ideal ninepeptides (HBx1,HBx2,HBx3,and HBx4) were selected as candidate peptides. Using flow cytometry,the fluorescence index of both control and experimental groups were detected and the 4 ninepeptides were evaluated with T2 binding assay and DC50 assay. Results: The ninepeptides VLCLRPVGA (HBx1),CLFKDWEEL (HBx2),VLHKRTLGL (HBx3) and HLSLRGLPV (HBx4) were selected as candidate targets. Among the 4 candidate peptides,HBx2 showed higher HLAA0201 affinity and HBx2,HBx4 showed better stability.Conclusion: Our study indicates that CLFKDWEEL might be a potential HLAA0201 restricted CTL epitope from hepatitis B virus X protein; further study is needed for verification of its immunity in vivo．