检测牛奶中结合珠蛋白的电流型免疫传感器的研制  

Development of an amperometric immunosensor for detection of haptoglobin in cow milk

作　　者：丁守强[1] 刘彬[2] 谭勋[1] 潘韬[1] 

机构地区：[1]浙江大学动物科学学院/农业部动物疫病病原学与免疫控制重点开放实验室,浙江杭州310029 [2]全国畜牧总站,北京100125

出　　处：《南京农业大学学报》2011年第5期112-116,共5页Journal of Nanjing Agricultural University

基　　金：浙江省科技厅项目(2007C32025)

摘　　要：为构建可定量检测牛奶中结合珠蛋白(Hp)含量的电流型免疫传感器,以三电极系统中的金电极为工作电极,采用层层自组装技术将L-半胱氨酸、纳米金颗粒和抗牛Hp抗体依次组装在工作电极上,制成免疫传感器。制成的免疫传感器对Hp的线性检测范围为15～100 mg.L-1,最低检测限为0.63 mg.L-1,并具有较好的重现性和稳定性。分别采用该传感器和ELISA检测试剂盒对隐性乳房炎患牛乳样中的Hp含量进行检测的结果表明,两种检测结果间无显著差异(P>0.05)。结论:制备的免疫传感器可定量检测牛奶中的结合珠蛋白(Hp)。Increased haptoglobin（Hp）concentration in milk is one of the indictors of subclinical mastitis in dairy cows.The present study was conducted to develop an amperometric immunosensor for quantitative detection of Hp in cow milk using three-electrode-system as basic apparatus.The gold electrode of the electrochemical system was defined to be the working electrode of immunosensor.L-cysteine,gold nanoparticles（GNP）and anti-bovine Hp antibody were fabricated in sequence onto the surface of the working electrode by layer-by-layer self-assembly technique.The immunosensor was of good reproducibility and stability,with detectable linear range between 15 and 100 mg·L-1 and a limit of detection（LOD）by 0.63 mg·L-1.The concentration of Hp in milk from cows with subclinical mastitis was detected using immunosensor and commercial ELISA kit,respectively,and agreement between the results obtained by the two methods was satisfactory（P0.05）.The immunosensor can be used to quantitatively detect the Hp in milk,thereby providing a promising method for diagnosis of subclinical mastitis in dairy cows.

关 键 词：牛奶 结合珠蛋白 免疫传感器 

分 类 号：S854.43[农业科学—临床兽医学]