基于金标抗体的电化学免疫传感器检测米糠油中黄曲霉毒素B1  

Detection of aflatoxin B1 in rice bran oil by electrochemical immunosensor based on gold labeled antibody

作　　者：郝小童 朱正伟 曲文莉 宫智勇 段烁 伍金娥 戴煌 杨庆 许琳[1,3,4] HAO Xiao-Tong;ZHU Zheng-Wei;QU Wen-Li;GONG Zhi-Yong;DUAN Shuo;WU Jin-E;DAI Huang;YANG Qing;XU Lin(College of Food Science and Engineering,Wuhan Polytechnic University,Wuhan 430023,China;Institute of Material Inspection of Hubei Bureau,National Food and Strategic Reserves Administration,Wuhan 430050,China;Key Laboratory for Deep Processing of Major Grain and Oil,Ministry of Education,Wuhan 430023,China;Hubei Key Laboratory for Processing and Transformation of Agricultural Products,Wuhan 430023,China)

机构地区：[1]武汉轻工大学食品科学与工程学院,武汉430023 [2]国家粮食和物资储备局湖北局物资检验所,武汉430050 [3]大宗粮油精深加工教育部重点实验室,武汉430023 [4]湖北省农产品加工与转化重点实验室,武汉430023

出　　处：《食品安全质量检测学报》2023年第20期252-259,共8页Journal of Food Safety and Quality

基　　金：武汉市知识创新专项(2022020801020399);湖北省重点研发计划项目(2021BCA113)。

摘　　要：目的构建金标抗体免疫探针并基于该探针建立一种电化学免疫传感器用于米糠油中黄曲霉毒素B1(aflatoxin B1,AFB1)的快速检测。方法首先在金电极(AuE)表面修饰抗原,加入制备好的金标抗体免疫探针和待测目标物,最后加入带有辣根过氧化物酶的二抗(IgG-HRP)。当目标物AFB_(1)存在时,AFB_(1)与抗原竞争结合金标抗体,使金电极表面结合的IgG-HRP减少,进而导致HRP催化过氧化氢将溶液中对苯二酚氧化成苯醌的电流信号下降,间接检测AFB1。采用GB 5009.22—2016《食品安全国家标准食品中黄曲霉毒素B族和G族的测定》对样品进行前处理,并优化了抗原用量、AuNP孵育时间等参数。结果该免疫传感器在抗原包被量和金标抗体用量为3μL且金标抗体与AFB1孵育80 min条件下具有最佳性能。该法线性范围为0.02~80.00 ng/mL,检出限为0.027 ng/mL。将该传感器应用于米糠油中AFB1检测,回收率在88.7%~108.0%。结论该传感器具有良好的特异性、重现性和稳定性,可用于米糠油中AFB1的快速检测。Objective To construct a gold labeled antibody immune probe and establish an electrochemical immunosensor based on this probe for the rapid detection of aflatoxin B_(1)(AFB_(1))in rice bran oil.Methods Firstly,the antigen was modified on the surface of gold electrode(AuE),and the prepared gold labeled antibody immunoprobe and the target were added.Finally,the second antibody with horseradish peroxidase(IgG-HRP)was added.When the target AFB_(1) existed,AFB1 competed with the antigen to bind the gold labeled antibody,which reduced the IgG-HRP bound on the surface of the gold electrode,and then led to the decrease of the current signal of HRP catalyzing hydrogen peroxide to oxidize hydroquinone into benzoquinone in the solution,thus detecting AFB_(1) indirectly.The samples were pretreated by GB 5009.22—2016 National food safety standards-Determination of aflatoxins B and G in food,and the parameters such as antigen dosage and AuNP incubation time were optimized.Results The immunosensor had the best performance when the amount of coating antigen and gold labeled antibody was 3μL,and the gold labeled antibody was incubated with AFB_(1) for 80 min.The linear range of this method was 0.02‒80.00 ng/mL,and the limit of detection was 0.027 ng/mL.The sensor was applied to detect AFB1 in rice bran oil,and the recovery rate was 88.7%‒108.0%.Conclusion The sensor demonstrates good specificity,reproducibility and stability,and can be used for the rapid detection of AFB1 in rice bran oil.

关 键 词：黄曲霉毒素B1 电化学免疫传感器 金标抗体 米糠油 快速检测 

分 类 号：TS225.19[轻工技术与工程—粮食、油脂及植物蛋白工程] O657.1[轻工技术与工程—食品科学与工程]