胶体金免疫层析法快速检测韭菜中腐霉利残留

【目的】建立韭菜中腐霉利残留的胶体金免疫层析快速检测方法。【方法】通过合成两种不同结构的腐霉利半抗原,并分别与载体蛋白耦联制得免疫原和包被原,免疫小鼠获得腐霉利单克隆抗体。基于竞争性免疫抑制原理,建立可视化胶体金免疫层析法。【结果】检测韭菜中腐霉利残留,检测限为5 mg/kg,满足国家标准规定的最大残留限量;对实际样本的检测结果与液相色谱-质谱法检测结果一致。【结论】本研究建立的方法具有简便、快速、准确等优点,可实现对韭菜中腐霉利残留超标的高通量快速筛选。

Preparation of an anti-diethylstilbestrol monoclonal antibody and development of an indirect competitive ELISA to detect diethylstilbestrol in biological samples

Based on the preparation of an anti-diethylstilbestrol(DES) monoclonal antibody,a simple and convenient indirect competitive enzyme-linked immunosorbent assay(ELISA) method for DES detection has been developed.The monoclonal antibody demonstrated high sensitivity to DES with an IC50 value of 275 pg mL-1 and detection limit(LOD) of 90 pg mL-1.The specificity of the assay was studied by measuring cross-reactivity of the antibody with structurally related compounds of ethinyl estradiol(<7%),estrone(<0.1%),estriol(<0.1%),and diethylstilbestrol benzoate(<0.1%).Chicken,fish,shrimp,urine and bile spiked with different concentration of DES were detected by the developed method,and the recovery rates were greater than 79.5%.Intra-and inter-assay variations were about 6%.This method exhibited high stability with a coefficient of variation less than 10% in buffer and in real samples.The LODs in fish/shrimp,liver,feed and urine spiked with DES were 600,600,4800 and 600 pg mL-1,respectively.These results confirmed that the antibody to DES was successfully produced and could be used to establish ELISA methods for DES detection in food producing animals.

基于胶体金免疫层析技术的赤霉素快筛试纸条的研制

研制快速检测豆芽中赤霉素的快筛试纸条。采用免疫层析技术,以胶体金为标记物制备赤霉素单克隆抗体-胶体金偶联物,以竞争抑制模式制备赤霉素胶体金免疫层析试纸条。该试纸条对豆芽中赤霉素的检测限为100 μg/kg,灵敏度为98%,特异性为97%,假阴性率为2%,假阳性率为3%。该方法操作简便,具有较高的灵敏度和特异性,可广泛应用于豆芽中赤霉素残留的现场筛查和检测。