磺胺嘧啶残留阻断ELISA试剂盒的研制及鉴定

Development and Trait Identification of Rapid Test Blocking ELISA Kit for Sulfadiazine Residues

基于1株分泌抗磺胺嘧啶(sulfadiazine,SD)高亲和力的SD单克隆抗体(monoclonal antibody,mAb),应用ELISA试验原理研制SD残留快速检测阻断ELISA试剂盒(SD-Kit),并对其特性进行测定。SD-Kit的标准曲线呈典型的S型,相关系数R2=0.9914,符合4参数logit曲线拟合,线性检测范围为0.02～12.92μg/L,灵敏度为0.08μg/L,半数抑制浓度(IC50)为0.54μg/L,检测限为0.08μg/L;鲜奶样和猪尿样的平均添加回收率分别为90.8%、95.8%,平均批内和批间变异系数均低于10%;基质对SD-Kit的检测结果影响不大;SD-Kit与磺胺甲基嘧啶的交叉反应率为2.7%,与其他磺胺类药几乎没有反应性;试剂盒在4℃可保存6个月。

A blocking ELISA test kit for detection of sulfadiazine（SD）was developed based on SD monoclonal antibodies（mAb）.The calibration curve of the SD-Kit with standard SD inhibitor was typical sigmoid curve fitted to the four parameter logistic equation with the linear detection of 0.02 to 12.92μg/L（R2=0.9914）,the sensitivity of 0.08μg/L,the IC50 of 0.54μg/L and the detection limit of 0.08μg/L.The recoveries of SD spiked in milk were 90.8%,in pig urine were 95.8%.The precision and accuracy of the assay as determined by inter-assay and intra-assay coefficient variation was both below 10%.The SD-Kit generally had 2.7% cross-reactivity towards sulfamerazine and little or no cross-reactivity towards other sulfonamides.The dilution solution of SD had no effect on results of SD-Kit.The validity of SD-Kit in 4℃ was above six months.