[关键词]
[摘要]
为检测鲤鱼中苯并咪唑类药物残留,降低其对人体造成的危害,本文基于单克隆抗体之上建立胶体金试纸条检测方法。首先以2-甲氧基羰基氨基-3H-苯并咪唑-5-羧酸为半抗原,用活化酯法偶联蛋白质制备免疫原,免疫小鼠,进行细胞融合制备单克隆抗体,而后用合成的胶体金标记单克隆抗体制备胶体金试纸条。通过间接竞争ELSIA法(ic-ELISA)测得阿苯达唑、甲苯咪唑、阿苯达唑亚砜、阿苯达唑砜、芬苯达唑、氟苯咪唑、奥芬达唑的半抑制浓度(IC50)分别为0.44、0.16、3.47、5.62、0.62、0.10、5.77 ng/mL。胶体金试纸条在鲤鱼中的阿苯达唑、甲苯咪唑、阿苯达唑亚砜、阿苯达唑砜、芬苯达唑、氟苯咪唑、奥芬达唑的检测限分别为4、2、50、100、10、1、100 ng/g,检测时间为15 min。因此,该方法灵敏度高、成本低、速度快且无需仪器辅助,适宜现场大量鲤鱼样本的快速检测。
[Key word]
[Abstract]
In order to reduce its harm to human, a colloidal gold immunochromatographic strip was established based on monoclonal antibody (mAb) to detect the residue of benzimidazoles in carp. Firstly, the immunogen was prepared using 2-(methoxycarbonylamino)- 3H-benzimidazole-5-carboxylic acid as hapten and coupling protein by activated ester method, mice were immunized and mAb was prepared by cell fusion. Then A colloidal gold immunochromatographic strip was prepared by mAb labelled with synthetic colloidal gold. The 50% inhibitory concentrations (IC50) of albendazole, mebendazole, albendazole s-oxide, albendazole sulfone, fenbendazole flubendazole, oxfendazole were 0.44, 0.16, 3.47, 5.62, 0.62, 0.10, 5.77 ng/mL by ic-ELISA, respectively. The limit of detection (LOD) of albendazole, mebendazole, albendazole s-oxide, albendazole sulfone, fenbendazole, flubendazole, oxfendazole were 4, 2, 50, 100, 10, 1, 100 ng/g in carp samples, respectively. Therefore, the developed method was highly sensitive, rapid, low-cost, free-instrument and applicable for on-site detection of a large numbers of carp samples.
[中图分类号]
[基金项目]
国家重点研发计划项目(2017YFC1600801)