对硫磷直接免疫测定方法的初步研究

A Direct Competitive Enzyme-linked Immunosorbent Assay for the Determination of Parathion

通过制备对硫磷的特异性抗体,建立了对硫磷的直接竞 争酶联免疫吸附分析法。先将对硫磷苯环上的硝基还原为氨 基,然后采用重氮化法将氨基-对硫磷与牛血清白蛋白相偶 联。对合成的抗原进行紫外可见扫描,确证偶联成功。用免 疫抗原免疫新西兰大耳白兔,制得对硫磷抗血清,测得其效价 为1.2×104,与其他类似物的交叉反应为甲基对硫磷: 1.22%,毒死蜱:0.09%,杀螟松:0.06%,标准曲线的最佳检 测范围为7.2～720.0ng/mL。用此血清建立的ELISA方法 与GC方法的相关系数可达94%,用该方法检测蔬菜中添加 的对硫磷标准样品,平均回收率为107.77%。

Direct competitive Enzyme linked immunosorbent assay (ELISA) was developed for the analysis of parathion. The nitro group in parathion was reduced to amino group. A diazotization reaction was employed to conjugate reduced parathion to large carrier protein. The antigen was identified by the method of ultraviolet and visible light scanning. The result demonstrated that the conjugates were synthesized successfully. The anti parathion serum was obtained after three months by using the immunogen in rabbits. The titer of antiserum was 1.2 10 4 , Parathion could be quantitated in the range 7.2~720.0 ng/ml. The cross reaction tests indicated that most analogues and the compounds structurally similar to parathion did not showed cross reaction to the antibodies. The ELISA and GC method has a 94% correlation coefficient. Average recovery of parathion from vegetables was107.77%.