应用FITC系统建立非均衡竞争总甲状腺素(TT4)化学发光法  

Construction of non-equilibrium competitive chemiluminesce immunoassay using FITC system for the detection of total tetraiodothyronine

作　　者：陈蕾[1] 孔海霞[2] 刘萍[2,3] 宋启超 田云霞 

机构地区：[1]中国康复研究中心北京博爱医院,北京100068 [2]重庆医科大学检验医学院,重庆400016 [3]博奥赛斯(天津)生物科技有限公司,天津300162

出　　处：《国际检验医学杂志》2012年第6期663-665,共3页International Journal of Laboratory Medicine

基　　金：天津市科技型中小企业技术创新专项资金项目(09ZXCXSH05300)

摘　　要：目的建立可广泛应用于总甲状腺素(TT4)检测的方法。方法制备辣根过氧化物酶(HRP)标记兔抗人甲状腺素(T4)多克隆抗体及异硫氰酸荧光素(FITC)标记T4类似物。以抗FITC抗体包被发光板,FITC-T4类似物与抗FITC抗体结合形成固相抗原,血清标本经处理后使T4均呈游离状态,固相抗原与血清T4竞争结合抗T4抗体-HRP,建立化学发光法(CLIA)血清TT4检测系统(FITC系统),进行方法学评价,并与T4-牛血清γ球蛋白直接包被系统(非FITC系统)和罗氏公司Elec-sys2010系统进行比较。结果 FITC系统检测线性范围为0.1～320ng/mL,灵敏度为0.1ng/mL;批内变异系数小于5%,优于非FITC系统;与Elecsys2010系统检测结果具有良好的相关性(r=0.9169)。结论应用FITC系统建立的非均衡竞争TT4CLIA分析系统灵敏度高、特异性,可用于临床标本的检测。Objective To construct a method for the detection of total tetraiodothyronine（TT4）.Methods Anti-tetraiodothyronine（T4） polyclonal antibody was labeled with horse radish peroxidase（HRP）,and fluorescein isothiocyanate（FITC） was conjugated to T4-analogue.Microwell was coated by anti-FITC antibody,FITC-T4-analogue was combined with anti-FITC to form solid phase antigen.T4 in serum samples,treated to release combined T4,and solid phage antigen would combine to anti-T4-HRP competively（FITC system）.FITC system was evaluated and compared with non-FITC system,based on T4-bovine serum gamma globulin,and Roche Elecsys2010 system.Results The constructed FITC system was with linear range of 0.1-320 ng/mL,sensitivity of 0.1 ng/mL,and coefficients of variability within-run under 5%,which was better than non-FITC system.There was fine correlation between FITC system and Elecsys2010 system（r=0.916 9）.Conclusion Non-equilibrium competitive CLIA system based on FITC system was successfully constructed,with satisfying specificity and sensitivity,and could be applied for the detection of clinical samples.

关 键 词：异硫氰酸荧光素 非均衡竞争 甲状腺素 化学发光免疫分析 

分 类 号：R446.6[医药卫生—诊断学]