聚多巴胺侧流免疫层析法检测谷物中呕吐毒素

Detection of the deoxynivalenol in cereals by polydopamine-based lateral flow immunochromatography assay

中文关键词:  聚多巴胺  呕吐毒素  免疫学试纸条  检测

英文关键词:polydopamine  deoxynivalenol  lateral flow immunochromatography assay  detection

中文摘要:

      目的 制备一种基于聚多巴胺(polydopamine, PDA)纳米载体的免疫层析试纸条, 用于高灵敏检测谷物中的呕吐毒素(deoxynivalenol, DON)。方法 本研究通过多巴胺自氧化聚合制备PDA, 一步结合法制备PDA-单克隆抗体(monoclonal antibody, PDA-mAb)探针, 以呕吐毒素抗原为检测线(T线)、羊抗鼠二抗作为质控线(C线)建立PDA免疫层析试纸条。结果 该试纸条检测DON的消线值为6.0 ng/mL, 视觉检出限为0.9 ng/mL, IC50=1.13 ng/mL, 比荧光微球试纸条和胶体金试纸条分别高0.88倍和3.08倍, 且该试纸条稳定性与特异性良好。此外, 该试纸条已经成功用于玉米、小米和燕麦样品中DON的检测, 最低检出限分别为8.40、6.87、9.89 μg/kg, 回收率为80.10%~122.05%, 相对标准偏差均小于12.01%。结论 本研究建立的PDA免疫层析试纸条具有灵敏、准确、快速和简便等特点, 为谷物中真菌毒素的检测提供技术支撑。

英文摘要:

      Objective To prepare a polydopamine (PDA)-based lateral flow immunochromatography assay for the highly sensitive detection of deoxynivalenol (DON) in cereals. Methods In this paper, PDA was synthesized by dopamine self-oxidative polymerization, and the PDA labeled monoclonal antibody (PDA-mAb) probe was prepared by a one-step conjugation method. Subsequently, the PDA lateral flow immunochromatography assay was established using DON antigen as the test line (T line) and goat anti-mouse antibody as the control line (C line). Results The results showed that the cut-off value of the PDA lateral flow immunochromatography assay for detecting DON was 6.0 ng/mL, the visual limit of detection was 0.9 ng/mL, half maximal inhibitory concentration was 1.13 ng/mL, which were 0.88 times and 3.08 times higher than those of fluorescent microsphere test strip and colloidal gold test strip, respectively. Meanwhile, the PDA lateral flow immunochromatography assay possessed well stability and specificity. In addition, the proposed lateral flow immunochromatography assay has been successfully used for the detection of DON in corn, millet, and oats samples, with minimum detection limits of 8.40, 6.87, and 9.89 μg/kg, recoveries ranging from 80.10%–122.05%, and relative standard deviations less than 12.01%. Conclusion In summary, the PDA lateral flow immunochromatography assay established in this work is sensitive, accurate, rapid and simple, and provides technical support for the detection of mycotoxins in cereals.