大白菜中拟除虫菊酯类农药残留半定量ELISA免疫检测方法的建立

Development of a semi-quantitative ELISA for detection of pyrethroid pesticides residue in Chinese cabbage

为建立适合蔬菜中拟除虫菊酯类农药的半定量ELISA免疫检测方法,以实验室自制的针对拟除虫菊酯类农药的广谱性抗体为材料,以间苯氧基苯甲酸(PBA)为半抗原,分别用活性酯法、混合酸酐法和6-氨基己酸法偶联鸡卵清蛋白(OVA)制备3种包被抗原,开展了半定量酶联免疫分析法(ELISA)免疫检测试验。结果表明,采用6-氨基己酸法合成包被抗原效果最优;检测时无需将抗原抗体过夜提前预混,抗原抗体反应时间以45 min较佳;无净化步骤的大白菜基质对检测结果影响较小,该方法对63个大白菜样品的假阳性检出率和假阴性检出率分别为9.3%和2.7%,表明该方法具备对大量大白菜样品快速初筛的应用潜力。

The broad spectrum antibody against a group of pyrethroid insecticides was used as the material, 3-phe-noxybenzoic acid （PBA） was used as generic hapten, and three coating antigens were synthesized by activated estermethod, mixed anhydride method and 6-aminocaproic acid method, for detection of pyrethroid pesticide residues in Chi-nese cabbage by semi-quantititative ELISA（OVA）. The 6-aminocaproic acid method the best coating antigen for identifi-cation of pyrethroid pesticides without premixing theantigen and antibody. The optimum reaction time forantigen and antibody was 45 min. The semi-quantitativeELISA developed showed false positive rate and false neg-ative rate of 9.3% and 2.7% for 63 Chinese cabbagesamples, indicating that this method is a potential meas-ure for fast identification of pyrethroid pesticides residuein Chinese cabbage.