hTK1质量放大压电石英晶体免疫传感器研究  

Amplified mass quartz crystal immunosensor for human thymidine kinase I

作　　者：丁克祥[1] 吴朝阳[2] 郑永晨[3] 王攀[4] 周颖 曹国彬[6] 胡燕祝[7] 吕沅津 

机构地区：[1]南方医科大学衰老生物医学研究中心,广东广州510515 [2]湖南大学化学化工学院 [3]吉林大学第二医院中心实验室 [4]武汉理工大学复杂系统研究中心 [5]湖北省中山医院 [6]广州医学院第二附属医院 [7]北京盛泽伟业科技发展公司 [8]上海海京征生物科技发展公司

出　　处：《中国老年学杂志》2005年第9期1064-1067,共4页Chinese Journal of Gerontology

基　　金：国家863计划项目(2002AA217051);国家科技部2002年度国际科技合作重点项目〔国科发外字(2003)26号项目序号No:67〕;上海市科委2004年度科研计划项目(No:044119668)

摘　　要：目的研制高灵敏度的质量放大压电石英晶体免疫传感器用于人血清中癌症标志物胸苷激酶1(hTK1) 的检测.方法将半胱氨自组装膜用戊二醛活化,共价固定hTK1抗原,通过和待测抗原竞争与一定浓度的酶标抗体反应,应用4-氯-1萘酚和H2O2质量放大体系,测得样品中待测抗原的浓度.结果 hTK1 的浓度在 0.1～10 ng/ml 范围内,与传感器频率的变化之间呈良好的线性关系.应用此方法对患有不同癌症病人血清进行测定,发现正常血清与癌症血清的频移值存在显着差异.结论基于质量放大原理的压电石英晶体免疫传感器可定量测定血清中的 ng 水平的 hTK1 抗原,有望用于癌症的早期诊断和临床检测.Objective To develop a new amplified mass piezoelectric quartz crystal immunosensor with high sensitivity for the determination of serum human thymidine kinase Ⅰ （hTKⅠ）. Methods The monomolecular films of cysteamine were first self-assembled onto the gold electrode surfaces of quartz crystals, and hTKⅠ antigens were covalently immobilized via difunctional reagent, glutaraldehyde. The immobilized hTKⅠ antigens and free hTKⅠantigens in samples competitively reacted with the horseradish peroxidase labeled anti-hTK1 antibodies, and free hTKⅠ antigens in samples were then indirectly determined through a 4-chloro-1-naphthol / H2O2 mass amplification system. Results There was a good linearity between frequency shifts and hTK1 concentrations in the range of 0.1 - 10 ng/ml. Moreover, several cancer serum samples were detected to find a significant difference in frequency response between cancer serum samples and normal serum. Conclusions The amplified mass piezoelectric quartz crystal immunosensors can quantitatively detect hTK1 at nanogram level,and their applications in clinic diagnosis of cancers can be expected.

关 键 词：人血清中胸苷激酶1 压电石英晶体 质量放大 免疫传感器 肿瘤早期诊断 

分 类 号：Q789[生物学—分子生物学]