三唑磷的免疫分析技术研究

Studies on the Immunoassay for Triazophos

将合成的三唑磷半抗原采用活性酯法分别与牛血清白蛋白和卵清蛋白共价偶联制备突出三唑磷分子结构特征的人工抗原与包被抗原.以人工抗原免疫新西兰白兔获得抗血清,采用硫酸铵分步盐析和DEAE 纤维素反相吸附法从抗血清中分离纯化对三唑磷具特异性亲合力的抗体,以辣根过氧化物酶采用混合酸酐法标记半抗原.在此基础上,首次成功建立了对三唑磷具高特异性的间接竞争、包被抗体直接竞争酶联免疫吸附分析(ELISA)技术.在优化条件下,三唑磷测定的线性浓度范围为0.001～1.0 mg/L,检出限0.11 μg/L,其他类似结构的常用有机磷酸酯类杀虫剂和苯唑醇不干扰三唑磷的测定.

The synthesized hapten of triazophos was covalently conjugated to bovine serum albumin and ovalbumin respectively to prepare artificial antigen and coating antigen. The artificial antigen was used as immunogen to immune New Zealand rabbits to prepare antiserum, from which the polyclonal antibodies with high affinity to triazophos was separated by the method of salting out and purified by the method of reversed phase adsorption of diethylaminoethyl （DEAE）-cellulose. The hapten was conjugated to horseradish peroxidase to prepare enzyme tagged hapten by means of mixed anhydride method. Based on these, the indirect competitive and antibody-coating direct competitive enzyme-linked immunosorbent assays （ELISA） were developed successfully. Under optimized conditions, the linear concentrations ranged from 0. 001 mg/L to 1.0 mg/L and the detection limit was 0. 11 μg/L for the determination of triazophos. Some analogues of triazophos, such as common used organic phosphate pesticides chlorpyrifos, parathion, didn＇t interfere with the analysis of triazophos