基于侧流免疫层析技术制备胶体金试纸条检测莲子中黄曲霉毒素B_1的研究

Determination of aflatoxin B_1 in lotus seeds by colloidal gold test strip based on lateral flow immune chromatography technique

目的基于侧流免疫层析技术制备一种适用于快速检测莲子中黄曲霉毒素B1(AFB1)的试纸条。方法利用柠檬酸三钠还原法制备胶体金溶液,金颗粒标记单克隆抗体制成金标偶联物作为结合垫,特异性抗原(AFB1-BSA)和羊抗鼠Ig G二抗分别作为检测线和控制线。结果经组装测试,试纸条的灵敏度为2.5ng·m L-1,检测时间在10 min以内。在检测的23份莲子样品中,6份样品有AFB1,阳性样品经UFLC-MS/MS确证,试纸条假阳性和假阴性率为0。结论所制备的试纸条检测快速、操作简便、结果准确,适合大批量莲子中AFB1的现场检测。

Objective To develop t he colloidal gold test strip based on the lateral flow immune chromatography technique for rapid detection of aflatoxin B1 in lotus seeds. Methods Firstly, the colloidal gold solution was prepared by sodium citrate reduction reaction. Then, the colloidal gold test strip was constructed based on the conjugation of gold particles with monoclonal antibody as the combined pad. The specific antigen of AFB1-BSA served as the text l ine and goat anti-mouse Ig G antibody as the control line, which were immobilized on the nitrocellulose filter membrane to construct the colloidal gold test strip. Results The detection limit of the constructed test strip was 2.5 ng·m L- 1 and all the lotus seed samples were analyzed within 10 min. In 23 batches of samples, 6 samples were detected AFB1. These positive samples were confirmed by UFLC-MS/MS. There were no false-positive and negative results.Conclusion The constructed colloidal gold test strip is simple, quick and accurate for detecting AFB1 in lo tus seeds.