荧光免疫定量法即时检测谷物中黄曲霉毒素B1  

Point-of-care testing of aflatoxins B1 in cereals by fluorescence immunoquantitative assay

作　　者：苗银萍 付燕峰 赵光升 瞿超杰 娄亚坤 李宁 任宝红 许洪超 赵林萍 MIAO Yin-Ping;FU Yan-Feng;ZHAO Guang-Sheng;QU Chao-Jie;LOU Ya-Kun;LI Ning;REN Bao-Hong;XU Hong-Chao;ZHAO Lin-Ping(Zhengzhou Zhongdao Biotechnology Co.,Ltd.,Zhengzhou 450000,China;State Key Laboratory of Market Regulation(Food Safety Rapid Testing and Smart Supervision Technology),Zhengzhou 450000,China;Henan Insitute of Food and Salt Industry Inspection Technology,Zhengzhou 450000,China)

机构地区：[1]郑州中道生物技术有限公司,郑州450000 [2]国家市场监管重点实验室(食品安全快速检测与智慧监管技术),郑州450000 [3]河南省食品和盐业检验技术研究院,郑州450000

出　　处：《食品安全质量检测学报》2023年第15期129-138,共10页Journal of Food Safety and Quality

基　　金：河南省市场监督管理局科技计划项目(2022sj26)。

摘　　要：目的建立荧光免疫定量法即时检测谷物中黄曲霉毒素B1的含量。方法以荧光微球为标记物,采用2,4二硝基苯酚独立质控体系和竞争法检测原理,构建谷物中黄曲霉毒素B1荧光免疫定量即时检测方法。评价其准确度、重复性、特异性、稳定性及与仪器确证方法符合度。结果该方法最适条件为:pH 7.8硼酸盐缓冲液活化、pH 7.5磷酸缓冲液偶联、微球抗体质量比为5:4、抗原抗体反应质量比为50:1。试纸条检出限为0.299~0.997μg/kg,定量限为0.544~2.663μg/kg。代表性样本准确度为92.20%~111.80%,重复性为3.2%~10.6%。除与黄曲霉毒素B2有交叉反应(6.1%),与其他类似物无交叉反应(<0.1%)。该法检测结果与超高效液相色谱法结果一致性好,相关系数(r^(2))为0.997。结论该方法准确、快速、灵敏、便捷,适合于现场即时检测谷物中黄曲霉毒素B1的含量。Objective To establish a method for the point-of-care testing of aflatoxins B1 in cereals by fluorescence immunoquantitative assay.Methods Using fluorescent microspheres as markers,point-of-care testing of aflatoxins B1 in cereals by fluorescence immunoquantitative assay was established.It adopted 2,4-dinitrophenol independent quality control system,and was based on the detection principle of competition method.The accuracy,repeatability,specificity,stability and conformity with the instrument confirmation method were evaluated.Results The optimum conditions for the method were as follows:pH 7.8 borate buffer activation,pH 7.5 phosphate buffer coupling,microsphere antibody mass ratio was 5:4,antigen antibody mass ratio was 50:1.The limits of detection were 0.299‒0.997μg/kg,and the limits of quantitation was 0.544‒2.663μg/kg.The accuracy of the representative samples ranged from 92.20%to 111.80%,and the repeatability ranged from 3.2%to 10.6%.Except for cross-reaction with aflatoxin B2(6.1%),there was no cross-reaction with other analogenates(<0.1%).The correlation coefficient(r^(2))was 0.997 comparing these 2 methods,the consistency was good.Conclusion The method is accurate,rapid,sensitive and convenient,and is suitable for the in-situ point-of-care testing of aflatoxins B1 in cereals.

关 键 词：谷物 黄曲霉毒素B1 荧光免疫 定量 即时检测 

分 类 号：TS210.7[轻工技术与工程—粮食、油脂及植物蛋白工程]