应用FITC系统建立非均衡竞争游离雌三醇(E3)化学发光法  

Construction of non-equilibrium competitive chemiluminesce immunoassay using FITC system for the detection of free estriol

作　　者：曾建明[1] 雷鹏[2] 孔海霞[2] 宋启超 刘萍[2,3] 

机构地区：[1]广东省中医院大学城医院检验科,广州510006 [2]重庆医科大学检验医学院,400016 [3]博奥赛斯(天津)生物科技有限公司,300162

出　　处：《国际检验医学杂志》2012年第15期1806-1808,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：目的建立可以广泛应用于游离雌三醇(E3)检测的方法。方法制备辣根过氧化物酶(HRP)标记兔抗人多克隆抗体及异硫氰酸荧光素(FITC)标记雌三醇类似物。以抗FITC抗体包被发光板,FITC-E3类似物与抗FITC抗体结合形成固相抗原,并与血清中的freeE3竞争结合抗E3抗体-HRP,建立化学发光血清freeE3检测系统(FITC系统),进行方法学评价,并与E3-牛血清白蛋白直接包被系统(非FITC系统)和西门子ADVIACentaur?XP系统进行比较。结果 FITC系统检测线性范围为0.1～40ng/mL,灵敏度为0.1ng/mL;批内变异系数小于5%优于非FITC系统;与西门子ADVIACentaur?XP系统检测结果有良好的相关性(r=0.9657)。结论应用FITC系统建立的非均衡竞争freeE3化学发光免疫分析系统灵敏度高、特异性强,可用于临床标本的检测。Objective To construct a method for the detection of free estriol（E3）.Methods Anti-estriol polyclonal antibody was labeled with horse radish peroxidase（HRP）,and fluorescein isothiocyanate（FITC） was conjugated to estriol analogue.Microwell was coated by anti-FITC antibody,FITC-estriol analogue was combined with anti-FITC to form solid phase antigen,and solid phage antigen and estriol in serum samples would combine to anti-E3-HRP competitively（FITC system）.FITC system was evaluated and compared with non-FITC system,based on E3-bovine serum albumin,and Siemens ADVIA Centaur XP system.Results The constructed FITC system was with linear range of 0.1-40 ng/mL,sensitivity of 0.1 ng/mL and coefficients of variability among and between batch under 5%,which was better than non-FITC system.Compared with Siemens ADVIA Centaur XP system,the correlation coefficient was 0.9657,and with no statistical difference of detection results（P0.5）.Conclusion The non-equilibrium competitive chemiluminesce immunoassay based on FITC system was successfully constructed,with satisfying precision and sensitivity,and could be applied for the detection of clinical samples.

关 键 词：游离雌三醇 异硫氰酸荧光素 非均衡竞争 化学发光免疫分析 

分 类 号：R446.6[医药卫生—诊断学]