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应用FITC系统建立非均衡竞争睾酮(T)化学发光法 被引量:1

Construction of non-equilibrium competitive chemiluminesce immunoassay using FITC system for the detection of testosterone
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摘要 目的建立可广泛应用于睾酮(T)检测的方法。方法制备辣根过氧化物酶(HRP)标记兔抗人T多克隆抗体及异硫氰酸荧光素(FITC)标记T-类似物。以抗FITC抗体包被发光板,FITC-T类似物与抗FITC抗体结合形成固相抗原,固相抗原与血清中T竞争结合抗T抗体-HRP,建立化学发光法(CLIA)血清T检测系统(FITC系统),进行方法学评价,并与直接用T-牛血清清蛋白包被系统(非FITC系统)和罗氏公司Elecsys2010系统进行比较。结果 FITC系统检测线性范围为0.05~20ng/mL,灵敏度为0.05ng/mL;批内、批间变异系数均小于5%,优于非FITC系统;与Elecsys2010系统检测结果的相关系数为0.9732,测定结果差异无统计学意义(P>0.05)。结论成功建立基于FITC系统的非均衡竞争T检测CLIA系统,精密度、灵敏度等指标均符合临床要求,可用于临床标本检测。 Objective To construct a method for the detection of testosterone(T).Methods anti-T polyclonal antibody was labeled with horse radish peroxidase(HRP),and luorescein isothiocyanate(FITC) was conjugated to T-analogue.Microwell was coated by anti-FITC antibody,FITC-T-analogue was combined with anti-FITC to form solid phase antigen,and solid phage antigen and T in sample would combine to anti-T-HRP competively(FITC system).FITC system was evaluated and compared with non-FITC system,based on T-bovine serum albumin,and Roche Elecsys2010 system.Results The constructed FITC system was with linear range of 0.05-20 ng/mL,sensitivity of 0.05 ng/mL,and coefficients of variability among and between batch under 5%,which was better than non-FITC system.Compared with Elecsys2010 system,the correlation coefficient was 0.973 2,and with no statistical difference of detection resusts(P0.05).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.
出处 《国际检验医学杂志》 CAS 2012年第6期668-670,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)
关键词 异硫氰酸荧光素 非均衡竞争 睾酮 化学发光免疫分析 fluorescein isothiocyanate non-equilibrium competition testosterone chemiluminescence immunoassay
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