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1、2期曾得意等：精喹禾灵酶联免疫吸咐分析（ELISA）研究449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200000。经酶联免疫吸附
2、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。关键词：精喹禾灵；半抗原；多克隆抗体；IC-ELISAStudyonELISAforQuizalofop-p-ethylZENGDe-yi1,2,WANGMing-hua1,SHI
3、Hai-yan1,LIBo1(1CollegeofPlantProtect,NanjingAgriculturalUniversity,Nanjing210095;2FineChemicalsResearchandDevelopmentCenter,GuizhouUniversity,Guiyang550025)Abstract:【Objective】Thepurposeofthestudyistoobtainspecialantibodyanddevelopcompetitiveindirectenzyme-linkedimmunosorbentassay(ciE
4、LISA)forquizalofop-p-ethyl.【Method】Quizalofop-p-ethylwashydrolyzedonthealkaliconditiontosynthesizehapten,quizalopacid,whichwascovalentlyconjugatedwiththecarrierproteinsbovineserumalbumin(BSA)asimmunogenandwithovalbumin(OVA)ascoatingantigen,respectively,andthecouplingratioofHatpen-BSAwa
5、s29.23andthecouplingratioofHatpen-OVAwas7.87.Polyclonalantibodyagainstquizalofop-p-ethylwasgeneratedbyimmunizingrabbitswithHatpen-BSA.【Result】Theantiserumwithhightiterthatwas200000.Theresultsshowedthattheantibodyhadhighaffinitytoquizalofop-p-ethyltestedbyenzymelinkedimmunsosorbentassay
6、(ELISA),theIC50forquizalofop-p-ethylwas0.03495μg·ml-1andthelowestdetectionlimitwas0.002μg·ml-1,theworkingrangewasassignedtoconcentrations0.002-0.5µg·ml-1forquizalofop-p-ethyl.Furthermore,themethodhadhighspecificityforitdidnotcross-reactwithotherstructure-relatedcompounds,andwhenwatersa
7、mplesfortifiedwithquizalofop-p-ethylwereanalyzed,recoverieswereintherangeof89.02%-108.88%.【Conclusion】Specialantibodywasproducedsuccessfullyandacompetitiveindirectenzyme-linkedimmunosorbentassay(ciELISA)wasdevelopedfortheanalysisofquizalofop-p-ethylresidueinwater.Keywords:Quizalofop-
