Objective To Identify B-cell epitopes on monkey B virus envelope protein gD. Methods Base on bioinformatics software, secondary structure, hydrophilicity, surface accessibility, antigenic index and flexibility of monkey B virus envelope protein gD was analysed and some potential peptide epitopes were forecasted. Then, the interactions between synthetic peptides and BV positive sera were detected by Enzyme-linked immunosorbent assay (ELISA). At last, The sensitivity and specificity of synthetic peptides was evaluated used 20 samples of Standard sera by ELISA. Result Seven epitopes were forecasted by Bioinformatics analysis. Four synthetic peptides, sequence as ⁴⁶LPPLEQKTD⁵⁴, ¹⁰⁶RGAPEATRSDA¹¹⁶, ²⁹¹PELAPEERGTSRTPGD³⁰⁶ and ³⁶¹AVYLVRRRGR³⁷⁰could be reacted with positive sera pool. The sensitivity of 4 synthetic peptides changed form 40% to 70% and the specificity were 100% for 4 synthetic peptides. Conclusion There are at least four linear epitope on B virus gD protein.