拟除虫菊酯类农药多残留酶联免疫分析方法的条件优化  

Optimization of Multi-residue Enzyme-linked Immunoassay for Pyrethroid Pesticides

作　　者：吴元元[1] 金朵[1] 付骋宇 祁志军[1,3] 

机构地区：[1]西北农林科技大学农药研究所,陕西杨凌712100 [2]陕西出入境检验检疫局,西安710068 [3]植物源农药研究与开发陕西省重点实验室,陕西杨凌712100

出　　处：《农药》2017年第7期510-514,共5页Agrochemicals

基　　金：国家自然科学基金项目(30900955);国家检验检疫总局科技项目(2014IK016)

摘　　要：[目的]优化拟除虫菊酯类农药多残留检测条件,建立适用于多种菊酯类农药的酶联免疫分析方法。[方法]优化多残留酶联免疫检测方法,建立半抗原对单链抗体的标准抑制曲线。对7种拟除虫菊酯类农药进行交叉反应,计算交叉反应率。[结果]最佳检测条件:包被抗原以质量浓度5 mg/L于37℃包被微孔2 h、单链抗体稀释640倍、稀释液中甲醇体积分数为10%、竞争时间为80 min、HRP/Anti-His酶标二抗稀释质量浓度为1/7000 g/L、显色20 min。该方法对氯菊酯、氯氰菊酯和溴氰菊酯有较强交叉反应,交叉率分别为62.31%、43.43%、36.93%。[结论]成功建立可检测多种拟除虫菊酯类农药的酶联免疫分析方法,为免疫分析试剂盒的研发奠定基础。[Aims] The aims were to optimize the detection conditions for pyrethroid pesticide residues and establish an enzyme-linked immunoassay method for a variety ofpyrethroid pesticides. [Methods] The multi-residue enzyme-linked immunoassay conditions for pyrethroid pesticides were optimized. The standard inhibition curve for hapten to single chain antibody was established. The cross-reactivity of seven pyrethroid pesticides was calculated. [Results] The optimal detection conditions were as follows： the concentration of coated antigen was 5 mg/L, 37℃ for 2 h, single chain Fv （scFv） was diluted by 640 times, the volume fraction of methanol in the dilution was 10%, the compete time was 80 rain, the concentration of HRP/Anti-His secondary enzyme-antibody was 1/7000 g/L, coloration for 20 min. This scFv has a strong cross reactivity to permethrin, cypermethrin and deltamethrin, the cross rates were 62.31, 43.43 and 36.93%, respectively. [Conclusions] The method of enzyme-linked immtmoassay for detecting pyrethroid pesticides was successfully established, which laid the foundation for the development of immunoassay kit.

关 键 词：拟除虫菊酯 半抗原 多残留 酶联免疫分析 条件优化 交叉反应 

分 类 号：TQ450.26[化学工程—农药化工]