Abstract: BALB/c mice were immunized with streptomycin-bovine serum albumin and hybridoma lines that secrete monoclonal antibody against streptomycin(SM mAb) were filtered with cell fusion. A competitive ELISA test kit for detection of streptomycin（SM-Kit）was developed with anti-SM monoclonal antibody（SM mAb） and its sensitivity，veracity，specificity and the effect of matrix were tested, respectively. At the same time, the urine of the swine was detected with this ELISA-Kit. The results indicated that the SM-Kit has little or no crossreactivity with other antibiotics besides of dihydrostreptomycin；the linear detection was 1.0 to 128.0 μg/L（R2＝0.925 1），the sensitivity was 0.45 μg/L，the IC50 was 7.76 μg/L and the detection limit was 1 μg/L. The recoveries of SM spiked in pig urine was 84.1%. The matrix has no effect on the results of the SM-Kit. The SM-Kit possessed rapidity, sensitivity, specificity and briefness and was proved useful in the rapid test of SM residues in animal.