动物源性食品中泰乐菌素残留ELISA试剂盒的研制

Development of Test ELISA Kit for Tylosin Residues in Animal-derived Food

关键词:  泰乐菌素  单克隆抗体  竞争ELISA

Key words:  Tylosin  Monoclonal antibodies  Competitive ELISA

摘要:

本文采用杂交瘤技术，筛选并克隆出能够稳定分泌抗泰乐菌素(TYL)单抗的杂交瘤细胞株，免疫BALB/c小白鼠制备抗泰乐菌素单克隆抗体，建立泰乐菌素竞争ELISA检测试剂盒。结果表明，Logit/Log拟合标准曲线为y= -1.84x+2.02，相关系数r=0.9944，线性检测范围为1.5~121.5ug/L ，半数抑制浓度(IC50)为10.3ug/L，灵敏度为1.5ug/L，检测限为1.5~3.0ug/L；肌肉、肝脏、蜂蜜的添加回收率分别为64%-99%、61%-102%、60.5%-95%；平均批内和批间变异系数均小于10%；泰乐菌素与替米卡星（TIL）的交叉反应为40%，与其他类抗生素无交叉反应；此泰乐菌素试剂盒在4℃可保存6个月以上。本文建立的竞争ELISA检测方法可用于动物源性食品中泰乐菌素残留的定量检测。

Abstract:

An competitive ELISA kit for detection of Tylosin (TYL) residues was developed using a TYL monoclonal antibodies, which was produced by hybridoma technology via immunizing BALB/c mouses. Results showed that the kit’s Logit/Log calibration curve was y= -1.84x+2.02 (correlation coefficient, r=0.9944), linear detection ranging from 1.5ug/L to 121.5ug/L, with half inhibitory concentration (IC50) 10.3ug/L, sensitivity 1.5ug/L, detection limit ranging from 1.5ug/L to 3.0ug/L.The added coefficient of recovery for TYL in muscle, liver, and honey is respectively 64%-99%、61%-102%、60.5%-95%,with coefficient of variation(CV%) lower than 10%, cross-reactivity between TYL and TIL 40%,and no cross-reactivity between TYL and other large ring lactone antibiotics .Moreover, the ELISA kit could be stored at 4℃ for more than six months ,and there is a conclusion that the ELISA kit could be applied to the quantitative detection for TYL residues in animal-derived foods.