食品中违法添加奥沙普嗪的荧光免疫层析方法研究  

Fluorescent immunochromatographic assay for screening oxaprozin as an illegal adulterant in food

作　　者：陈华龙 黄婉琼 张敏滢 刘萌 姜丽君 郭杰标[2] CHEN Hua-long;HUANG Wan-Qiong;ZHANG Min-ying;LIU Meng;JIANG Li-jun;GUO Jie-biao(Shaoguan Institute for Food and Drug Control, Shaoguan Guangdong 512026;Shaoguan College, Shaoguan Guangdong 512005)

机构地区：[1]韶关市食品药品检验所,广东韶关512026 [2]韶关学院,广东韶关512005

出　　处：《中南药学》2018年第4期518-521,共4页Central South Pharmacy

基　　金：广东省食品药品检验检测技术创新项目(No.2015ZX09);广东大学生科技创新培育专项资金(攀登计划专项资金pdjh2017b0457);全国大学生创新训练计划立项(No.201410576009)

摘　　要：目的建立一种筛查食品中违法添加的奥沙普嗪荧光微球免疫层析法(FICA)。方法以改良活泼酯法将奥沙普嗪连接到牛血白蛋白和卵清白蛋白,合成免疫抗原和检测抗原,免疫家兔获得针对抗奥沙普嗪抗体;以抗体标记的荧光微球(40 nm),在检测线和质控线发生免疫反应,分别形成荧光检测信号,构建荧光免疫层析方法。结果在ELISA体系检测奥沙普嗪的IC50值为13.2 ng·m L-1,与8种相关药物交叉反应率都小于0.1%,优化FICA试剂条对奥沙普嗪的最低检测限为2.0 ng·m L-1。优化的FICA试剂对50个市售样品的检测结果,与HPLC验证结果吻合。结论本方法灵敏、特异、简便、快速,是筛查食品中违法添加奥沙普嗪的有效工具。Objective To develop a fluorescent immunochromatographic assay （FICA） to screen oxaprozin as an illegal adulterant in food. Methods Immunogen and testing antigen were prepared by conjugating oxaprozin to BSA and OVA with modified active ester mechanism, respectively. Antibodies against oxaprozin were raised by immunization in rabbits with immunogen. Fluorescent micro-spheres （40 nm） labeled with antibodies formed fluorescence signals on the testing line and control line through immunological reaction, respectively. Results The IC50 value for oxaprozin was 13.2 ng· mL-1 in the ELISA. The cross-reactivities with 8 associate drugs were lower than 0.1%. A consequent FICA showed the detection limit of 2.0 ng · mL-1 for oxaprozin. Current FICA showed good agreements with HPLC in detecting oxaprozin in 50 food samples from the local market. Conclusion FICA is suitable in screening the illegal adulterant such as oxaprozin in food due to its high sensitivity and specificity.

关 键 词：奥沙普嗪 人工抗原 免疫微球 荧光免疫层析方法 

分 类 号：R392.11[医药卫生—免疫学] R446.61[医药卫生—基础医学]