应用FITC系统建立雌二醇直接竞争化学发光免疫分析方法  

Establishment of Direct Competitive Chemiluminescence Immunoassay for Estradiol with FITC System

作　　者：潘国华 PAN Guo-hua(Beijing North Institute of Biological Technology,Beijing100076,China)

机构地区：[1]北京北方生物技术研究所有限公司,北京100076

出　　处：《标记免疫分析与临床》2019年第2期351-354,360,共5页Labeled Immunoassays and Clinical Medicine

摘　　要：目的利用异硫氰酸荧光素(fluorescein isothiocyanate,FITC)系统建立人血清雌二醇(estradiol,E_2)直接竞争化学发光免疫分析方法。方法采用兔抗-FITC抗体、FITC-抗-E_2两步包被制备预包被板,采用直接竞争法原理建立E_2化学发光免疫分析检测体系,并对FITC-anti-E_2包被浓度、酶标抗原稀释比例、缓冲体系、反应时间等各种影响因素进行优化。结果本系统线性范围为30～3000 pg/mL,空白检测限4.8pg/mL,定量检测限30 pg/mL,分析内精密度5.6%,批间精密性8.7%,平均回收率95.9%,与雌三醇、睾酮、孕酮均无明显交叉反应,与贝克曼全自动化学发光试剂同时测定164份样本,二者测定结果的相关系数r为0.979。结论成功建立了雌二醇直接竞争化学发光免疫分析方法 ,该方法性能指标优秀,具有一定的临床应用价值。Objective A direct competitive chemiluminescence immunoassay for human serum estradiol (E 2 )was established using the fluorescein isothiocyanate (FITC)system. Methods Two-step coating of rabbit anti-FITC antibody and FITC-anti-E 2 were used to prepare the pre-clad plate. The direct competitive method was applied,investigated and optimized,to establish the new CLIA detection system for E 2. The E 2 chemiluminescence immunoassay detection system was established by the principle of direct competition,considering a variety of factors such as dilution ratio of enzyme-labeled antigen,the concentration of FITC-anti-E 2 coating,buffer system and incubate time. Results The linear detection range was 30~3000 pg/mL in this system,with the parameters as:the limit of the detection 4.8pg/mL,the limit of quantitative detection 30 pg/mL,the precision within the analysis 5.6%,the inter-assay precision 8.7%,the recovery rate for accuracy 95.9%. There were no cross reactions with estriol ,testosterone and progesterone. The simultaneous determination of 164 samples with Beckman′s automated chemiluminescence reagents yielded a correlation coefficient of 0.979. Conclusion The CLIA system for detecting E 2 has been established,which has an excellent performance indicators and can be widely used in the clinical testing.

关 键 词：荧光素 雌二醇 化学发光 

分 类 号：R446.6[医药卫生—诊断学]