基于群青蓝纳米粒子的免疫层析法快速检测牛奶中的黄曲霉毒素M1

Rapid detection of aflatoxin M1 in milk by immunochromatographic assay based on ultramarine blue nanoparticles

中文关键词:  免疫层析法  群青蓝纳米粒子  黄曲霉毒素M1

英文关键词:immunochromatographic assay  ultramarine blue nanoparticles  aflatoxin M1

中文摘要:

      目的 建立一种基于群青蓝纳米粒子的免疫层析法(immunochromatographic assay, ICA)快速检测牛奶中的黄曲霉毒素M1 (aflatoxin M1, AFM1)的方法。方法 通过离心筛选出合适粒径的群青蓝纳米粒子, 经羧基化修饰后与AFM1单克隆抗体通过碳二亚胺法共价偶联, 制备免疫探针, 用于AFM1检测。实验结果通过肉眼观察并结合Image J软件进行定性定量分析, 并对所建立方法的灵敏度、特异性进行评价。结果 经条件优化后, 该方法定性视觉检出限为0.30 ng/mL, 定量限为0.051 ng/mL, 与其他真菌毒素无交叉反应, 具有良好的特异性。结论 该方法可应用于牛奶样品中AFM1检测, 样品检测结果符合国家限量标准, 同时操作简单、检测快速、易于判断, 适用于现场快速初步筛查。

英文摘要:

      Objective To establish a method for the rapid detection of aflatoxin M1 (AFM1) in milk by immunochromatographic assay (ICA) based on ultramarine blue nanoparticles. Methods Ultramarine blue nanoparticles with suitable particle size were screened out by centrifugation, modified by carboxylation, and covalently coupled with AFM1 monoclonal antibody by carbodiimide method to prepare immunoprobes for AFM1 detection. Results can be observed by naked-eye and combined with Image J software for qualitative and quantitative analysis. The sensitivity and specificity of the ICA were evaluated. Result After optimization, the ICA had the visual limit of detection and limit of quantitation were 0.30 ng/mL and 0.051 ng/mL, respectively. Meanwhile, the ICA showed satisfactory specificity with no cross activity with other mycotoxin. Conclusion The ICA can be applied to detect AFM1 in milk, which meet the national standard. Besides, it had advantage of simple operation, rapid detection and easy judgment. Therefore, ultramarine blue nanoparticles based immunochromatographic assay was suitable for rapid preliminary screening in the field.