精喹禾灵酶联免疫吸咐分析ELISA研究-矿业科学学报.DOC

精喹禾灵酶联免疫吸咐分析（ELISA）研究 曾得意1,2，王鸣华1，施海燕1，李 波1 （1南京农业大学植物保护学院，南京 210095；2 贵州大学精细化工研究开发中心，贵阳 550025） 摘要：【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200 000。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0.03495 μg·ml-1，最低检测浓度（IC10）为0.002 μg·ml-1，检测范围为0.002～0.5 μg·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。 关键词：精喹禾灵；半抗原；多克隆抗体；IC-ELISA Study on ELISA for Quizalofop-p-ethyl ZENG De-yi1,2, WANG Ming-hua1, SHI Hai-yan1, LI Bo1 (1College of Plant Protect, Nanjing Agricultural University, Nanjing 210095; 2Fine Chemicals Research and Development Center, Guizhou University, Guiyang 550025) Abstract: 【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】Spec