黄曲霉素M1间接竞争ELISA的建立及其在牛奶中的应用  

Development of an Indirect Competitive ELISA for Rapid Analysis of Aflatoxin M1 in Milk

作　　者：陈曦[1] 孟萌[2] Sergei A Eremin 许迎辉 尹永梅[2] 郗日沫[2] 

机构地区：[1]天津科技大学生物工程学院,工业发酵微生物教育部重点实验室,天津市工业微生物重点实验室,天津300457 [2]南开大学药学院,南开大学药物化学生物学国家重点实验室,天津市分子药物研究重点实验室,天津300071 [3]莫斯科大学化学学院,莫斯科119991 [4]河北省石家庄市餐饮化保执法大队,石家庄050000

出　　处：《科学技术与工程》2013年第19期5455-5458,共4页Science Technology and Engineering

基　　金：天津市自然科学基金(11ZCGHHZ01200;12JCQNJC08900);中央高校基本科研业务费(65011751)资助

摘　　要：利用高特异性单克隆抗体,建立黄曲霉素M1的间接竞争ELISA检测方法。棋盘法优化了包被抗原及酶标抗体的最佳稀释倍数。包被抗原和酶标抗体的最佳工作浓度分别为1∶15000和1∶2000。该方法的线性范围为(0.1—8.1)ng·mL-1。除了与黄曲霉素B1的交叉反应为35%外,与黄曲霉素B2、黄曲霉素G1及黄曲霉素G2的交叉反应均小于1%。在牛奶实际样品检测的回收率均大于70%,而且检测过程在10min以内即可完成。该方法灵敏度高,特异性强,而且操作简单,适合多样本的快速筛查,为黄曲霉素M1的高效检测提供了一个良好的选择。An indirect competitive ELISA for Aflatoxin M1 was developed based on anti-AFM1 monoclonal an- tibody. Firstly, the working concentration of coated antigen and enzyme labeled antibody was optimized and their dilution ratios were selected as 1 ： 15 000 and 1 ： 2 000 respectively. The linear range of the proposed ELISA was in 0. 1--8. 1 （ng·mL-l）. The cross-reaetivities to AFB2, AFG1 and AFG2 were all below 1% , while that of AFB1 was 35%. The recoveries in milk samples were all higher than 70% and the detection could be finished within 10 min. It can be concluded that this method for AFM1 analysis was sensitive, specific, simple and rapid which exhibited a potentiality of screening a large number of samples on site. It also provided an option for effective determination of AFM1.

关 键 词：黄曲霉素M1 间接竞争ELISA 交叉反应 牛奶 

分 类 号：R446.9[医药卫生—诊断学]