Rapid diagnosis of foot-and-mouth disease virus (FMDV) is significant for the prevention of the FMD outbreak. In this study, we developed polyvinylidene difluoride (PVDF) supported polydiacetylene (PDA) immunosensor to detect FMDV, where VP1 polyclonal antibody was directly conjugated as a virus binding module without linker length. The optimal constituent ratio of the key platform was determined as 10,12-pentacosadiynoyl VP1 polyclonal antibody: dimyristoyl phosphatidyl choline: 10,12-pentacosadiynoic acid 1:4:5. With this composition, photo-polymerized colloids were investigated, and PVDF solid-state sensing was evaluated for the easy and portable platform. The detailed analyses were performed using UV-vis spectroscopy, DLS, FT-IR, TEM, and SEM. Resultingly, the blue-to-red color transition and fluorescent signal were successfully observed in the presence of FMDV antigen. This work provides a rapid and simple methodology for the FMDV detection.