Based on conventional enzyme linked immunoassay (ELISA) , a convenient, rapid and accurate one-step sulfonamides (SAs) residue chemiluminescent enzyme immunoassay (CLEIA) detection method was established through obtaining monoclonal antibodies from immunizing mice which was injected with SAs-BSA coupling protein, and optimizing test parameters such as antibody concentration, reaction time. The results showed that the best detection range of this method was 1.0~81.0ng/mL, the IC50 was 3.047ng/mL, the detection limit was 0.74ng/mL. Compared with conventional ELISA, the CLEIA method can not only increase the detection limit and sensitivity, but also shorten the reaction time by 60 minutes. CLEIA kit is more appropriate for rapid detection of the SAs residue in food.