动物组织中磺胺二甲嘧啶残留ELISA试剂盒研制

Development of ELISA Kit for Sulfamethazine Residues in Animal Tissues

采用重氮化法和戊二醛法,将磺胺二甲嘧啶分别与牛血清白蛋白和辣根过氧化物酶偶联制备了免疫原和酶标半抗原,免疫兔获得了特异性抗体,成功建立了相关动物产品中磺胺二甲嘧啶残留ELISA定量检测方法及商品化试剂盒,并对试剂盒的灵敏度、准确度、精密度和稳定性进行了研究.试剂盒检测线性范围为 62.5～0.54 ng/mL.在待测样品中各添加 500、200、100、50 ng/g SMZ,测试的回收率平均为89.0%～134.8%;试剂盒测定结果与色谱的平均符合率99.8%～126.0%;对比定性测试 15 份色谱检测为阴性的样品,均未出现假阳性.试剂盒存放在 37 ℃ 10 d和 2～8 ℃ 5 个月,质量稳定.

BSA-SMZ and HRP-SMZ conjugates were prepared using sulfamethazine coupled to BSA and horseradish peroxidase respectively. Specific antibodies against sulfamethazine were prepared using BSA-SMZ as antigens for the immunization of rabbits. Enzyme-Linked Immunosorbent Assay （ELISA） method and kit has been developed for quantitative examination BSA-SMZ residues in animal tissues. The sensitivity, accuracy, precision and stability of the kit were tested. The linear range determined was 0,54- 62, 5 ng/mL. The recovery rate of sulfamethazine in tested samples adding 500,200, 100, and 50 ng/g ranged from 89,0 % to 134.8 %. No false-positive result in negative samples was detected by HPLC. The quality of the kit stored at 2-8℃ for five months and 37 ℃ for ten days was stable.