精喹禾灵酶联免疫吸咐分析(ELISA)研究

Study on ELISA for Quizalofop-p-ethyl

【目的】制备精喹禾灵的特异性抗体并建立精喹禾灵的间接竞争ELISA测定方法。【方法】精喹禾灵在碱性条件下水解合成了半抗原精喹禾灵酸,并与载体蛋白BSA和OVA偶联,分别合成了免疫抗原和包被抗原,偶联比分别为29.23和7.87。将Hapten-BSA免疫兔子获得多克隆抗体。【结果】抗血清的效价为200000。经酶联免疫吸附反应分析(ELISA)测定,精喹禾灵的抑制中浓度(IC50)为0.03495μg·ml-1,最低检测浓度(IC10)为0.002μg·ml-1,检测范围为0.002～0.5μg·ml-1。其与结构相似的物质几乎没有交叉反应,表明该方法具有很强的特异性,且在水样中的添加回收率为89.02%～108.88%。【结论】成功获得精喹禾灵特异性抗体并建立了水样中精喹禾灵农药残留的ELISA测定方法。

【Objective】The purpose of the study is to obtain special antibody and develop competitive indirect enzyme-linked immunosorbent assay （ciELISA） for quizalofop-p-ethyl.【Method】Quizalofop-p-ethyl was hydrolyzed on the alkali condition to synthesize hapten, quizalop acid, which was covalently conjugated with the carrier proteins bovine serum albumin （BSA） as immunogen and with ovalbumin （OVA） as coating antigen, respectively, and the coupling ratio of Hatpen-BSA was 29.23 and the coupling ratio of Hatpen-OVA was 7.87. Polyclonal antibody against quizalofop-p-ethyl was generated by immunizing rabbits with Hatpen-BSA.【Result】The antiserum with high titer that was 200 000. The results showed that the antibody had high affinity to quizalofop-p-ethyl tested by enzyme linked immunsosorbent assay （ELISA）, the IC50 for quizalofop-p-ethyl was 0.03495 μg·ml-1 and the lowest detection limit was 0.002 μg·ml-1, the working range was assigned to concentrations 0.002-0.5 μg·ml-1 for quizalofop-p-ethyl. Furthermore, the method had high specificity for it did not cross-react with other structure-related compounds, and when water samples fortified with quizalofop-p-ethyl were analyzed, recoveries were in the range of 89.02%-108.88%.【Conclusion】Special antibody was produced successfully and a competitive indirect enzyme-linked immunosorbent assay （ciELISA） was developed for the analysis of quizalofop-p-ethyl residue in water.