一种新型多元均相时间分辨荧光免疫分析方法的建立  

A novel homogeneous time-resolved multiplexed fluoro-immunoassays

作　　者：陈振华[1] 陈玫君[1] 侯经远[1] 郑洁纯 吴英松[1] 刘天才[1] 

机构地区：[1]南方医科大学生物技术学院,广东广州510515 [2]南方医科大学公共卫生与热带医学学院,广东广州510515

出　　处：《湖北大学学报（自然科学版）》2013年第4期474-479,共6页Journal of Hubei University：Natural Science

基　　金：国家自然科学基金青年科学基金(30901382);国家自然科学基金面上项目(81271931);高等学校博士学科点专项科研基金新教师基金(20094433120008);广东省自然科学基金(S2012010009547);广东省高等学校人才引进专项资金(2009);南方医科大学引进人才科研启动基金(2009)资助

摘　　要：制备出具有光学特异性的3种不同颜色量子点纳米微球和铽螯合物,分别与CEA、AFP和HBsAg的两株抗体偶联,采用双抗体夹心法模式,充分利用两株抗体与抗原的相互作用,通过采集不同量子点发出的荧光信号,根据其信号的强弱判定标记在量子点纳米微球上的抗体结合的抗原的量,从而实现多元待分析物的定性定量分析.结果表明,量子点的荧光信号与CEA、AFP和HBsAg抗原浓度呈线性关系,其函数分别为:lgY=2.791 73+0.915 84lgX(R=0.998 08)、lgY=3.695 43+0.456 44lgX(R=0.998 48)和lgY=4.128 98+0.409 45lgX(R=0.998 45),分析灵敏度分别为:0.44、0.24和0.02ng/mL.Three kinds of quantum dots nanoparticles and terbium chelates were, respectively, prepared with novel optical properties. They were conjugated with two monoclonal antibodies respectively. Then with double antibodies sandwich assay, the mechanism of time-resolved fluoresce resonance energy transfer was mainly investigated by interactions between two monoclonal antibodies and their corresponding antigens. Detemiined the amount of antigens captured by antibodies based on the fluorescence signal from different quantum dots, in order to achieve qualitative and quantitative analysis of multiplex objects. The result showed that the signals had linear correlation with the concemration of antigens and they could be described as the following models： lgY=2. 791 73 ＋0. 915 841gX（R=0. 998 08）, lgY=3. 695 43＋0. 456 441gX（R=0. 998 48） and lgY=4. 128 98＋0. 409 451gX （R^0. 998 45）, the analytical sensitivities were 0. 44, 0. 24, and 0. 02 ng/mL.

关 键 词：多元均相免疫分析 时间分辨荧光免疫分析 双抗体夹心法 荧光共振能量转移 量子点 

分 类 号：R446.1[医药卫生—诊断学]