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

中文关键词:  粮谷物  黄曲霉毒素B1  荧光免疫  定量  即时检测

英文关键词:cereals  aflatoxins B1  fluorescence immunity  quantitative  point-of-care testing

中文摘要:

      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0.299~0.997 μg/kg, 定量限为0.544~2.663 μg/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(r2)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。

英文摘要:

      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?0.997 μg/kg, and the limits of quantitation was 0.544?2.663 μg/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 (r2) 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.