摘要
目的建立可以广泛应用于孕酮(P)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)标记孕酮类似物。以抗FITC抗体包被发光板,FITC-P类似物与抗FITC抗体结合形成固相抗原,并与血清中的P竞争结合抗P抗体-HRP,建立化学发光血清P检测系统(FITC系统),进行方法学评价,并与P-牛血清白蛋白直接包被系统(非FITC系统)和罗氏Elecsys2010系统进行比较。结果 FITC系统检测线性范围为0.1～60ng/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系统检测结果有良好的相关性(r=0.9619)。结论应用FITC系统建立的非均衡竞争P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 progesterone（P）.Methods Anti-progesterone polyclonal antibody was labeled with horse radish peroxidase（HRP）,and fluorescein isothiocyanate（FITC） was conjugated to progesterone analogue.Microwell was coated by anti-FITC antibody,FITC-progesterone analogue was combined with anti-FITC to form solid phase antigen,and solid phage antigen and progesterone in serum samples would combine to anti-P-HRP competitively（FITC system）.FITC system was evaluated and compared with non-FITC system,based on P-bovine serum albumin,and Roche Elecsys 2010 system.Results The constructed FITC system was with linear range of 0.1-60 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 Elecsys 2010 system,the correlation coefficient was 0.961 9,and with no statistical difference of detection results（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15期1813-1814,1817,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
孕酮
异硫氰酸荧光素
非均衡竞争
化学发光免疫分析
Progesterone
Fluorescein isothiocyanate
Non-equilibrium competition
Chemiluminescence immunoassay
