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M_(1)  

Rapid detection of aflatoxin M_(1) in milk by immunochromatographic assay based on ultramarine blue nanoparticles

作　　者：范耀龙 陈怡怡 朱军莉[1] 陆海霞[1] FAN Yao-Long;CHEN Yi-Yi;ZHU Jun-Li;LU Hai-Xia(College of Food Science and Biotechnology,Zhejiang Gongshang University,Hangzhou 310018,China)

机构地区：[1]浙江工商大学食品与生物工程学院,杭州310018

出　　处：《食品安全质量检测学报》2022年第10期3221-3227,共7页Journal of Food Safety and Quality

基　　金：浙江省基础公益研究计划项目(LGN2OC200007、LGN22C200025)。

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M_(1)(aflatoxin M_(1),AFM_(1))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AFM_(1)单克隆抗体通过碳二亚胺法共价偶联,制备免疫探针,用于AFM_(1)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AFM_(1)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。Objective To establish a method for the rapid detection of aflatoxin M_(1)(AFM_(1))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 AFM_(1) monoclonal antibody by carbodiimide method to prepare immunoprobes for AFM_(1) 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 AFM_(1) 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.

关 键 词：免疫层析法 群青蓝纳米粒子 黄曲霉毒素M_(1) 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]