甲硝唑人工抗原的制备及其ELISA检测

Synthesis and Identification of Metronidazole Artificial Antigen and the Development of ELISA Method

采用碳二亚胺法将甲硝唑半抗原与牛血清白蛋白(BSA)连接制备人工免疫原,同样方法将其与卵清蛋白(OVA)连接制备人工包被原。经紫外扫描分析,两种方法合成的免疫原和包被原的结合比分别为8：1、2：1和6：1、2：1；动物免疫试验分析偶联物,小鼠抗体效价达1：5 000,与3种硝基咪唑类药物的交叉反应率均小于10%。表明制备的抗体可以用于甲硝唑残留的检测,同时为检测试剂盒的研制奠定了基础。

Two methods of metronidazole artificial hapten was reported in this paper. The immunogen, metronidazole-BSA, was synthesized by carbodimide method. The coating antigen, metronidazole-OVA, was synthesized by the same method. The couple rates of the immunogens and coating antigens are 8 ： 1,2 ： 1 and 6 ： 1,2 ： 1,respectively. It showed the titer of Balb/c mice was 1 ： 5 000,and the cross-reaction was lower than 10% comparing to the other three Nitromidazoles. The results demonstrated that the two artificial antigens were successfully synthesized, which provides a basis for next monoclonal antibody preparation and ELISA kit development.