氟乐灵抗体的制备及其酶联免疫分析方法的建立

该研究针对农(水)产品中氟乐灵农药残留的问题,建立了间接竞争酶联免疫分析方法,快速检测氟乐灵农药.利用3,5-二硝基-4-氯-三氟甲苯与仲胺侧链氨基反应合成半抗原2C,采用碳二亚胺法将半抗原2C与牛血清白蛋白/卵清蛋白偶联合成完全抗原.通过免疫新西兰大白兔获得多克隆抗体PcAb-2C,基于间接竞争酶联免疫分析方法原理对...

Preparation of Trifluralin Antibody and Development of Enzyme Linked Immunosorbent Assay

An indirect competitive enzyme linked immunosorbent assay (ic-ELISA) was developed to rapidly detect trifluralin pesticide in agricultural or aquatic products in this paper. Hapten 2C was synthesized by the reaction of 4-chloro-3,5-dinitrobenzotrifluoride and secondary amine side chain amino group. And then hapten 2C was coupled with BSA or OVA to synthesize complete antigens by carbodiimide method. After that, polyclonal antibody PcAb-2C was obtained by immunizing New Zealand rabbits. Based on the principle of indirect enzyme linked immunosorbent assay, seven factors including dilution ratio of coating antigen and antibody, coating temperature, antibody and standard diluent were systematically optimized, and finally the indirect competitive enzyme linked immunosorbent assay was developed. Under the optimized conditions, the IC50 of this developed ic-ELISA was 54.07 ng/mL, the detection limit was 7.22 ng/mL, and the linear range was 12.56~282.62 ng/mL. The cross reaction with 20 trifluralin analogues showed good specificity (The cross reaction rate of two drugs was more than 15%). The recoveries of soybeans, soybean oil and eels were 89.8%~106.4%, and the coefficients of variation were less than 10%. The developed method was verified by gas chromatography and the recoveries were 89%~108.2% with the coefficients of variation less than 10%. The results showed a good linear relationship (R2=0.99). The above data show that the established method in this study has the advantages of high sensitivity, high specificity and high accuracy, which is suitable for the high-throughput rapid detection of trifluralin pesticide residues in agricultural (aquatic) products.