Objective: To predict the B-cell linear epitopes of E6 and E7 proteins of human papillomavirus （HPV）16 and HPV18 and prepare rabbit polyclonal antibodies against E6 and E7 of HPV16 and HPV18. Methods：Bioinformatics software was used to analyze the amino acid sequences of E6 and E7 proteins of HPV16 and HPV18 to predict the B-cell linear epitopes. The peptides of HPV16 E6, HPV16 E7, HPV18 E6 and HPV18 E7 were designed and synthesized. The synthesized peptides were cross-linked with Keyhole Limpet Hemocyanin (KLH), which was used to immunize New Zealand white rabbits to prepare the polyclonal antibodies. Recombinant proteins HPV16 E6 E7 and HPV18 E6 E7 expressed in the prokaryotic system were used as antigens to detect the titer and specificity of rabbit polyclonal antibodies by indirect ELISA and Western blot. Results: The titers of HPV16 E6, HPV16 E7 and HPV18 E7 polyclonal antibodies were 1:2.56×105, and the titer of HPV18 E6 polyclonal antibody was 1:1.28×105. Western blot results showed that the antibodies could bind specifically to the recombinant proteins HPV16 E6 E7 and HPV18 E6 E7. Conclusion: The polyclonal antibodies against E6 and E7 of HPV16 and HPV18 were successfully prepared, which created the conditions for the subsequent experimental research on HPV infection or HPV vaccine development.