采用高灵敏度的化学发光酶免疫分析方法(CLEIA)研究测定了环境水体及人体尿液中17β-雌二醇(17β-estradiol)的含量。以FITC-anti-FITC体系为固相,生物素-亲和素体系作为信号放大体系,高灵敏度的3-(2′-螺旋金刚烷)-4-甲氧基-4-(3″-羟基)苯1,2-二氧杂环丁烷磷酸钠(AMPPD)-碱性磷酸酶(ALP)化学发光反应作为信号检测体系。优化分析条件后,测量的线性范围为2.5-2000ng/L;检出限为1.1ng/L;批内、批间分析误差均在10%以内,稀释回收率和加标回收率均在92%-112%之间。测得环境水样中雌二醇的含量在0.15-0.57ng/L范围内,而人体尿液中雌二醇含量在0·75-135μg/L范围内。
A highly sensitive chemiluminescent enzyme immunoassay (CLEIA) for the determination of the 17β-estradiol (E2 ) in river and human urine was developed by using the FITC-anti-FITC system as solid phase, the avidin-biotin system as signal amplification system, and the highly sensitive 4-methoxy-4-(3-phosphatephenyl) -spiro-( 1,2-dioxetane-3,2'-adamantane ) ( AMPPD ) -alkaline phosphatase ( ALP ) system as chemiluminescence detection. After optimizing the conditions of enzyme reaction, the linear range of E2 was in the range of 2.5 - 2000 ng/L, detection limit was 1.1 ng/L. The RSDs of intra- and inter-assay were both below 10%. The percent recoveries are in the range of 92% - 112% in human urine. This method has been successfully applied to detect E2 in river water and human urine. The concentration of E2 in river water was in the range of 0. 15 -0.57 ng/L, and in human urine was in the range of 0. 75 -135 μg/L.