黄曲霉毒素M1检测ELISA条件的优化及应用研究  

Optimization of ELISA Method Conditions for Detection of AFM1 and it’s Application

作　　者：裴世春[1] 郑丽娜[1] 唐彦君[1] 张园园[1] 何娜[1] 

机构地区：[1]黑龙江八一农垦大学食品学院,大庆163319

出　　处：《黑龙江八一农垦大学学报》2008年第5期57-60,共4页journal of heilongjiang bayi agricultural university

基　　金：黑龙江省留学归国人员基金(Lc05c11)

摘　　要：采用黄曲霉毒素M1与牛血清白蛋白偶联物抗原免疫Balb/C小鼠获得特异性抗体。利用此抗体,进行间接竞争ELISA法试验,确定了最佳抗原包被反应浓度为1.0μg·mL-1,最佳抗原包被条件为4℃过夜,一抗和酶标二抗IgG-HRP最佳工作浓度为1∶8000和1∶5000。建立间接竞争定量ELISA方法,该方法的最低检出限是0.08ng·mL-1,间接竞争ELISA50%抑制率浓度为0.62ng·mL-1,校正曲线的线性范围为0.04～5ng·mL-1。在此优化条件下制备的抗体检测黄曲霉毒素M1具有很高的特异性和灵敏性,具有实际应用价值。The specific antibody was obtained by immune Balb/C mice with Aflatoxin Ml-bovine serum albumin （BSA） conjugate （AFM1-BSA）. Using this antibody, a indirect competitive ELISA method for the detection of AFM1 was developed with the best concentration of antigen, antigen coating condition, monoclonal antibody and second antibody marked with HRP being of 1.0 μg·mL^-1, 4 ℃ overnight 1/8000 and 1/5000 respectively. The linear range for developed indirect competitive ELISA was 0.04-5 ng·mL^-1 AFM1. The limit of detection and the concentration required for 50% inhibition of binding were 0.08 and 0.62ng·mL^-1 AFM1. The antibody prepared by the optimal conditions was highly specific and sensitive on detecting AFM1, showing high practical application value.

关 键 词：黄曲霉毒素M1 酶联免疫吸附测定 单克隆抗体 

分 类 号：R446.61[医药卫生—诊断学]