S1 gene targeted libraries containing the major immunodominant region (1～2 367 bp) of PEDV spike glycoprotein were constructed by phage-display vectors based on filamentous phage strain fd-tet, in which the exogenous polypeptides were expressed in the N-terminal of gene Ⅲ coat protein. The S1 libraries were panned three times using the purified rabbit sera against PEDV. Three peptides, displayed on recombinant phages, showed strong binding affinity with the PEDV antisera, and were designated as S1P1 (248～280aa), S1P2 (442～499aa) and S1P3 (697～742aa) respectively. In ELISA and Western blot, the three peptides were all recognized by the PEDV antisera, but S1P3 showed strong binding activity. To further determine antigenicity of the three peptides, the antisera of S1P1-GST, S1P2-GST, S1P3-GST and their ligations GST fusion protein were prepared. In ELISA, S1P1-GST, S1P2-GST, S1P3-GST and S1P123-GST fusion proteins were able to induce the S1-specific antisera. The result of indirect immunofluorescence assay (IFA) demonstrated that the antisera induced by S1P2-GST, S1P3-GST and S1P123-GST fusion proteins possessed binding ability to the native S protein of PEDV cultured in Vero cells.