| 有机磷杀虫剂通用结构半抗原的设计及广谱特异性抗体的制备 |
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| 引用本文： | 刘贤进,颜春荣,刘媛,余向阳,张存政.有机磷杀虫剂通用结构半抗原的设计及广谱特异性抗体的制备[J].中国农业科学,2008,41(3):727-733. |
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| 作者姓名： | 刘贤进 颜春荣 刘媛 余向阳 张存政 |
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| 作者单位： | 江苏省农业科学院食品质量安全检测所,南京,210014 |
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| 基金项目： | 国家高技术研究发展计划(863计划)
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国家自然科学基金 |
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| 摘 要： | 【目的】制备针对有机磷农药的广谱特异性抗体，用于有机磷农药的免疫快速筛选检测。【方法】设计采用二乙基膦酸乙酸作为通用结构半抗原，用NHS-DCC法和EDC法合成两种人工抗原免疫新西兰大白兔制备抗血清。【结果】NHS-DCC法所制得抗血清的最高滴度为25 600， EDC法合成的抗原制得的抗血清滴度最高为6 400，均获得免疫应答。以二乙基膦酸乙酸为对象建立的间接竞争ELISA检测方法对二乙基膦酸乙酸最低检测浓度0.0035 μg?ml-1，抑制中浓度I50为0.182 μg?ml-1。经对12种常见磷酸酯类有机磷农药特异性检测反应试验，结果发现：所得抗体对毒死蜱、氧乐果、二嗪农、乙基对硫磷、丙溴磷、辛硫磷等农药有特异性反应，I50分别为0.12、0.21、0.24、0.78、0.97、3.8 μg?ml-1。【结论】该检测技术可用于毒死蜱、氧乐果、二嗪农、乙基对硫磷、丙溴磷等药剂的快速定性或半定量检测。
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| 关 键 词： | 有机磷杀虫剂 广谱性抗体 酶联免疫吸附试验（ELISA） |
| 收稿时间： | 2007-01-12 |
| 修稿时间： | 2007-04-29 |
Use of Generic Hapten in Production of Broad Specificity Anti-Organophosphorous Pesticides Antibody |
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| LIU Xian-jin,YAN Chun-rong,LIU Yuan,YU Xiang-yang,ZHANG Cun-zheng.Use of Generic Hapten in Production of Broad Specificity Anti-Organophosphorous Pesticides Antibody[J].Scientia Agricultura Sinica,2008,41(3):727-733. |
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| Authors: | LIU Xian-jin YAN Chun-rong LIU Yuan YU Xiang-yang ZHANG Cun-zheng |
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| Abstract: | Diethylphosphono acetic acid (DPA) was used as a generic hapten in order to generate broad specificity polycolonal antibodies against a group of organophosphorus pesticides. Six New Zealand white rabbits were immunized with immunogens sysnthesised by the mixed-anhydried method and EDC method, the titers of antiserum had reached 25600 and 6400, respectively. The CI-ELISA was conducted with the antiserum, the detection limit for DPA was 3.536ng/mL and the I50 was 0.182μg/mL. The antiserum also showed high affinities to chlorpyrifos, diazinon, omethoate, parathion-ethyl and profenofos, the I50 to these pesticides were 0.12，0.15，0.21，0.88, 0.97 and 2.5μg/mL. It is concluded that this assay is suitable screening tool for quantities and semi-quantities detection of the five organophosphorus pesticides. |
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| Keywords: | Phosphorous pesticides Broad specificity antibody Enzyme linked immunosorbent assay (ELISA |
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