基于可溶性虫卵抗原的血吸虫抗体检测免疫传感器  

The soluble egg antigen-based immunosensor for detection of Schistosome antibodies

作　　者：马静[1] 周卓晟[2] 徐勇[1] 刘红艳[2] 余枫华[1] 刘军[1] 杨春秀[1] 

机构地区：[1]湖北省宜昌市疾病预防控制中心,湖北宜昌443000 [2]华中科技大学同济医学院公共卫生学院教育部环境与健康重点实验室,武汉430030

出　　处：《中国卫生检验杂志》2011年第1期123-125,共3页Chinese Journal of Health Laboratory Technology

基　　金：宜昌市2009年医疗卫生科技攻关项目(A09310-42)

摘　　要：目的:将免疫法特异性和电化学法灵敏性相结合,建立一种高灵敏基于可溶性虫卵抗原的电化学免疫传感器,用于血吸虫抗体的检测。方法:巯基乙胺通过吸附作用在金电极表面形成的自组装单分子层(SAM),依次滴加2.5%戊二醛和血吸虫抗原(可溶性虫卵抗原,soluble egg antigen,SEA),用以捕获抗SEA抗体(兔多抗或人多抗),然后再与辣根过氧化物酶(HRP)标记的二抗形成免疫复合物。用循环伏安法检测还原峰的电流值。结果:在底物为添加了2μl 30%的双氧水及1.0 mmol/L对苯二酚的0.1 mol/L pH7.2的PBS缓冲液中,还原峰的电流值与抗SEA兔血清稀释度在10-3与10-6范围内成正比,相关系数为0.956。与抗SEA人血清稀释比在10-1与10-4范围内成正比,相关系数为0.988。在1∶40稀释度条件下,血吸虫病人与正常人血清的抗SEA还原峰的电流值相差7.4μA。结论:该方法可以用于血吸虫抗体的初步检测。Objective：Through the combination of specific immunoassay and sensitive electrochemical method,a highly sensitive electrochemical immunosensor for Schistosoma antibodies was established,which was based on the soluble egg antigen（SEA） of Schistosome.Methods： The self-assembled monolayer（SAM） was formed by mercaptoethylamine adsorbing on gold electrode surface,folowed the dropping of 2.5% glutaraldehyde and SEA to capture Schistosoma antibodies（rabbit or human polyclonal antibodies）.Then the immune complex formation was combined with secondary antibodies labeled by horseradish peroxidase（HRP）.The reduction peak current reduction peak current was detected by cyclic voltammetry.Results： The data showed that in the substrate of pH7.2 0.1 M PBS buffer which was amended by 1.0 mmol/L hydroquinone and the addtion of 2 μl 30% hydrogen peroxide.The reduction peak current value was proportional to the dilution of rabbit anti-schistosome serum from 10-3 to 10-6 and correlation coefficient is 0.956,while the correlation coefficient is 0.988 at the dilution of human anti-schistosome serum from 10-1 to 10-4.The difference value between the anti-SEA reduction peak current of a schistosomiasis patient and normal serum is 7.4 μA at 1∶40 dilution.Conclusion： The method can be used for the initial detection of schistosomiasis antibodies.

关 键 词：血吸虫抗原 SEA 酶免疫传感器 循环伏安 

分 类 号：R532.21[医药卫生—内科学]