In this study, the polyclonal antibody against furazolidone was produced with furazolidone coupling to protein carriers by a diazotization method and glutaraldehyde reaction, respectively. The antibody obtained showed good specificity toward furazolidone and various cross-reactivity toward nitrofurantoin, nitrofurazone, and furaltadone. Then, an indirect competitive enzyme-linked immunosorbent assay (ELISA) based on the antibody was first developed for multidetermination of four nitrofurans in animal feeds. The limit of detection (LOD) of the method was 0.2−2.1 ng/g depending on the component. After simple extraction, the fortified swine and broiler chicken feed samples were detected with recovery ranges of 75.9−86.4%. Results obtained from ELISA were confirmed by high-performance liquid chromatography (HPLC) with ultraviolet detection. Analysis of the unknown feed samples indicates that ELISA can be a practical tool for screening of nitrofurans in animal feeds before confirmation by HPLC.