目的 为粪便中其他小分子抗菌药物的酶联免疫分析方法的构建提供技术借鉴，实现猪粪中氧氟沙星禁用抗菌药物的养殖全过程管理。 方法 采用碳二亚胺法，将氧氟沙星（OFL）与载体蛋白牛血清白蛋白（BSA）和卵清白蛋白（OVA）偶联，制备出人工抗原OFL-BSA和OFL-OVA。使用OFL-OVA抗原制备氧氟沙星单克隆抗体，使用OFL-BSA作为包被抗原建立氧氟沙星间接竞争酶联免疫吸附（ELISA）方法。风干的猪粪样品经醋酸-醋酸钠溶液（pH约4.0）与甲醇的混合液（1∶3，V/V）提取及稀释后测定。 结果 建立的氧氟沙星间接竞争ELISA方法在标准溶液浓度范围（0~81 ng/mL）内线性良好（r>0.99），方法检测限为3.5 µg/kg。不同添加浓度的氧氟沙星回收率为70.8%~86.8%，变异系数为0.7%~4.7%。应用该法检测79份猪粪实际样品，检测结果与液相色谱-串联质谱法的分析结果具有良好的一致性。 结论 该方法准确、灵敏、快捷，可用于猪粪中氧氟沙星残留的快速分析。

Objectives A technical reference for establishing enzyme-linked immunosorbent assay (ELISA) methods for other small molecule antibiotics in feces was provided to realize the management in the entire process of farming to prohibit the use of antibiotics such as ofloxacin in pig manure. Methods The carbodiimide method was used to couple ofloxacin (OFL) with carrier proteins bovine serum albumin (BSA) and ovulin (OVA) to prepare artificial antigens OFL-BSA and OFL-OVA. OFL-OVA antigen was used to prepare monoclonal antibodies against ofloxacin. OFL-BSA was used as coated antigen to establish an indirect competitive ELISA method for ofloxacin. The air-dried pig manure samples were extracted and diluted with a mixture of acetic acid sodium acetate solution (pH approximately 4.0) and methanol (1∶3, V/V) for measurement. Results The indirect competitive ELISA method established for ofloxacin had good linearity (r>0.99) within the concentration of standard solution ranged from 0 ng/mL to 81 ng/mL, and the detection limit of the method was 3.5 µg/kg. The recovery rate of ofloxacin at different concentrations was ranged from 70.8% to 86.8%, with a coefficient of variation of 0.7% to 4.7%. The method was used to detect 79 actual samples of pig manure, and the results of detection were in good agreement with those of analysis with liquid chromatography tandem mass spectrometry. Conclusions This method established is accurate, sensitive, and fast, and can be used for rapidly analyzing residues of ofloxacin in pig manure.