In this study, 4-aminophthalic acid was used to synthesize hapten Dibutyl 4-aminophthalate by esterification reaction, and the synthesized hapten was coupled with keyhole limpet hemocyanin (KLH) by diazotization reaction to prepare the immune antigen and coupled with bovine serum albumin (BSA) to prepare the coating antigen. The New Zealand white rabbits were immunized to obtain the antiserum against dibutyl phthalate. An indirect competitive enzyme-linked immunosorbent assay (ELISA) for dibutyl phthalate was established using the purified antibodies. For the developed ELISA, the IC₅₀ was 40.68 μg/L and the limit of detection was 1.98 μg/L. The method was applied to detect liquor, milk and edible oil samples, and the recoveries ranged from 84.57% to 102.40%. There was a good correlation (R²=0.990 8) between the this ELISA and gas chromatography-mass spectrometry (GC-MS) for the detection of DBP in the spiked samples. The method established in this study can be applied to the rapid detection of dibutyl phthalate in food.