黄曲霉毒素B1免疫层析检测方法的研究

Study on colloidal gold immunochromatography assay for rapid detection of aflatoxin B1

以柠檬酸三钠为还原剂,制备了20nm胶体金颗粒,以此胶体金标记纯化的单抗4G6,喷于玻璃纤维上,AFB1蛋白偶联物和羊抗鼠IgG分别结合于硝酸纤维膜上,依次将样本垫、胶金垫、硝酸纤维膜和吸水纸组装,切割成胶体金试纸条。结果:组装的试纸条特异性好,与AF类似物无交叉反应,对AFB1标准品最低检测限为3μg/L,检测时间约为10min,批内和批间重复性为100%,假阳性率和假阴性率均为0。操作简便,成本很低,适于AFB1的现场快速检测。在制备了单克隆抗体的基础上,应用胶体金免疫层析技术,建立了食品中黄曲霉毒素B1(Aflatoxin B1,AFB1)快速检测方法。

Colloidal gold immunity chromotography technique was applied in this research. A rapid detection method for aflatoxin B1 in food was established. The method of trisodium citrate disoxidation was used to prepare colloidal gold particles which were used to mark AFB1 monoclonal antibodies and sprayed on the fiberglass. AFBl-protein and peroxidase-conjugated goat anti-mouse IgG combine on nitrocellulose independently,then sample tray, colloidal gold tray,nitrocellulose and bibulous paper were assembled to form colloidal gold test paper ti form the detection card. The result indicated the sensitivity of AFB1 fast detecting test paper was 45 μg/L;testing time was 10 min; the re- producibility between group inner and group exterior was 100 % ;false positive rate and false negative rate were all 0. It＇s simple and convenient for fast detecting of AFB1 in food on the spot.