中国农业科学 ›› 2008, Vol. 41 ›› Issue (3): 727-733 .doi: 10.3864/j.issn.0578-1752.2008.03.013

• 植物保护 • 上一篇    下一篇

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

刘贤进,颜春荣,刘 媛,余向阳,张存政   

  1. 江苏省农业科学院食品质量安全检测所
  • 收稿日期:2007-01-12 修回日期:1900-01-01 出版日期:2008-03-10 发布日期:2008-03-10

Use of Generic Hapten in the Production of Broad Specificity Anti-Organophosphorus Pesticides Antibody

  

  1. 江苏省农业科学院食品质量安全检测所
  • Received:2007-01-12 Revised:1900-01-01 Online:2008-03-10 Published:2008-03-10

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。

关键词: 有机磷杀虫剂, 广谱性抗体, 酶联免疫吸附试验(ELISA)

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.

Key words: Phosphorous pesticides, Broad specificity antibody, Enzyme linked immunosorbent assay (ELISA)