磺胺甲噁唑残留检测阻断ELISA试剂盒的研制

Development of Rapid Test Blocking ELISA Kit for Sulfamethoxazole Residues

在建立磺胺甲噁唑单克隆抗体（SMZmAb）杂交瘤细胞株和阻断ELISA方法的基础上，研制出SMZ残留快速检测试剂盒（SMZ—Kit），并对其性能进行了测定。结果表明，SMZ—Kit的标准曲线呈典型的S型，符合4参数logit曲线拟合，相关系数R^2=0，9901，检测范围为1．0～128．0μg／L，灵敏度为0．73μg／L，半数抑制浓度（IC50）为11．5μg／L，检测限为1．0μg／L；饲料样、牛奶样的平均添加回收率为83．9％，83．3％，平均批内和批间变异系数均〈15％；SMZ—Kit与磺胺嘧啶的交叉反应（CR％）为2．88％，与其他磺胺类药物无交叉反应；试剂盒在4℃可保存6个月。

A ELISA kit for detection of Sulfamethoxazole（SMZ-Kit） was developed with generating hybridoma lines that secrete SMZ monoelonal antibodies（SMZmAb） and the method of blocking ELISA, its traits were identified. The calibration curve of SMZ-Kit was typical sigmoid curve fitted to the four parameter logistic equation with the linear detection of 1.0 to 128.0μ g/L（R^2 = 0. 990 1 ）, the sensitivity of 0.73 μg/L, the IC50 of 11.5 μg/L and the detection limit of 1.0 μg/L. The recoveries of SMZ spiked in feed were 83.9 % and in milk were 83.3 % . The precision and accuracy of the assay as determined by inter-assay and intra-assay coefficient variation were below 15%. The SMZ-Kit had 2.88% cross-reactivity towards Suadiazine（SD）and little or no cross-reactivity towards other Sulfonamides. The validity of SMZ- Kit in 4℃ was above six months.