呋喃西林代谢物单克隆抗体及荧光免疫层析试纸条的制备

Preparation of Monoclonal Antibody and Fluorescence Immunochromatographic Test Strip for Detecting Furacilin Metabolites

为建立一种基于荧光免疫层析技术的呋喃西林代谢物的快速检测方法,采用活性酯法合成呋喃西林代谢物(氨基脲盐酸盐)(semicarbazide hydrochloride,SEM)人工抗原,并获得SEM的单克隆抗体,通过将量子点微球(quantumdot submicrobeads,QBs)与SEM偶联,得到QBs探针,制备荧光免疫层析试纸条。结果表明:SEM免疫荧光层析试纸条检测鱼肉组织的检测限为0.247?μg/L,不同添加质量浓度的回收率均在70%～120%之间,批内、批间变异系数均在15%以下,符合我国动物源食品中兽药残留检测方法相关指导文件的规定,且在2～8℃条件下可以保存6个月以上,稳定性良好。

In order to establish a rapid fluorescence immunochromatography method for the detection of semicarbazide hydrochloride(SEM)as a furacilin metabolite,SEM hapten was synthesized and conjugated to bovine serum albumin(BSA)and ovalbumin(OVA)by the active ester method,and the hapten-protein conjugates were employed to obtain anti-SEM monoclonal antibody.Meanwhile,SEM was conjugated to quantumdot microspheres,yielding a probe.Finally,a fluorescence immunochromatography test strip was prepared.The results showed that the detection limit of the test strip for fish meat was 0.247μg/L.The recoveries of SEM from blank fish samples spiked at different concentrations were between 70%and 120%,and within and between batches coefficients of variation(CV)were below 15%,which was in accordance with the requirements for the determination of veterinary drug residues in animal-derived foods.The test strip was stable at 2–8℃for up to 6 months.