基于流式荧光免疫法检测食品中米酵菌酸含量  

Detection of Bongkrekic Acid in Food by Flow Cytometry Fluorescence Immunoassay

作　　者：陆奕娜 粟有志 程铁辕 徐娟 伊雄海 刘俊 李志雄 李冠斯 LU Yi-na;SU You-zhi;CHENG Tie-yuan;XU Juan;YI Xiong-hai;LIU Jun;LI Zhi-xiong;LI Guan-si(Shantou Customs Technology Center,Shantou 515031,China;Yining Customs Technology Center,Yili 835000,China;Chengdu Customs Technology Center,Chengdu 610041,China;Guangzhou Customs Technology Center,Guangzhou 510623,China;Shanghai Customs Technical Center for Animal Plant and Food Inspection and Quarantine,Shanghai 200135,China;Guangzhou Darui Biotechnology Co.,Ltd,Guangzhou 510642,China)

机构地区：[1]汕头海关技术中心,广东汕头515031 [2]伊宁海关技术中心,新疆伊犁835000 [3]成都海关技术中心,四川成都610041 [4]广州海关技术中心,广东广州510623 [5]上海海关动植物与食品检验检疫技术中心,上海200135 [6]广州市达瑞生物技术股份有限公司,广东广州510642

出　　处：《分析测试学报》2025年第4期667-674,共8页Journal of Instrumental Analysis

基　　金：海关总署科研项目(2022HK108);汕头市科研项目(STKJ2023024)。

摘　　要：建立了一种定量检测米酵菌酸(BA)含量的流式荧光免疫法。首先通过合成BA-BSA完全抗原,免疫获得效价高的抗BA单克隆抗体。随后将BA-BSA合成抗原与藻红蛋白标记,并与样品中的BA竞争结合偶联有荧光微球的抗BA单克隆抗体,通过采用流式荧光仪检测微球表面的平均荧光强度,实现了样品中BA的定量测定。BA的线性范围为1.01~97.96μg/L,半数抑制浓度(IC_(50))为9.92μg/L,相关系数(r^(2))为0.9998,检出限为0.56μg/kg。加标回收率为90.4%~104%,相对标准偏差为5.0%~9.8%。方法特异性良好,与赭曲霉毒素A、黄曲霉素B1、黄曲霉素M1、伏马菌素B1、伏马菌素B2的交叉反应均<0.1%。实际样品测定结果与高效液相色谱-串联质谱法检测结果一致。所建立的流式荧光免疫法方法简单、快速、灵敏、准确,并可推广到各类食品等复杂基质中其他真菌毒素的分析和检测。In this study,a quantitative detection assay for bongkrekic acid(BA)based on flow cytometry fluorescence immunoassay was established.Synthesize BA-BSA complete antigen and obtain high titer monoclonal anti-BA antibody by immunization.BA-BSA labeled with phycoerythrin competes with BA in the sample for a limited number of antibody binding sites on the fluorescent microspheres.The average fluorescence intensity on the surface of the microspheres is detected by flow cytometry to achieve accurate quantitative determination of BA in the sample.The results showed that the assay achieved greatlinearity in the linear ranges from 1.01μg/L to 97.96μg/L,and the correlation coefficient(r^(2))was 0.9998.The limit of detection(LOD)was 0.56μg/kg.The recoveries in samples ranged from 90.4%to 104%,while the relative standard deviation was between 5.0%and 9.8%.The cross reactivities with ochratoxin A,aflatoxin B1,aflatoxin M1,fumonisin B1,and fumonisin B2 were all less than 0.1%.One positive sample was detected in 18 food samples,with a BA concentration of 6.5μg/kg,which was consistent with the results obtained by high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)instrument.The flow cytometry fluorescence immunoassay method established in this study is simple,rapid,sensitive,and accurate,and can be extended to the analysis and detection of other fungal toxins in complex matrices such as various foods.

关 键 词：流式荧光免疫法 米酵菌酸 抗原 抗体 食品安全 

分 类 号：O657.3[理学—分析化学] TS201.6[理学—化学]