中文摘要：

将抗兔IgG（即二抗）物理吸附于聚苯乙烯微孔板上作为通用固相，通过免疫反应制备固相抗体。采用辣根过氧化物酶催化鲁米诺-过氧化氢化学发光体系，建立了一种高通量、简便、快速的化学发光酶免疫分析方法用于临床测定人血清中的孕酮。对各种影响因素如免疫试剂的稀释度、发光底物选择、发光反应时间及温育条件等进行了考察和优化，最终选定的实验条件：孕酮抗体和HRP标记物的最佳稀释度分别为1：100000和1：15000；选用Ⅱ号发光底物，发光反应10min后测定；37℃水浴条件下温育1h。对建立的方法进行了评价。该方法的灵敏度为0．08μg／L；批内和批间相对标准偏差均在15％之内；低、中、高3个不同浓度值样品的平均回收率分别为101％、101％和94．4％（在87．8％～108％之间）。使用本方法和经典的放射免疫法同时对36份人血清样品进行测定，结果显示，本方法与放射免疫分析方法相关性良好，其相关系数为0．9502。

英文摘要：

A high-throughput, simple and rapid chemiluminescence enzyme immunoassay （CLEIA）was developed for the clinical determination of progesterone in human serum, and the luminol-hydrogen peroxide was used as chemiluminescence system catalyzed by horseradish peroxidase （HRP）. The solid phase of anti-progesterone antibody was prepared through the immunoreaction between anti-progesterone polyclonal antibody with donkey anti-rabbit IgG, i.e. second antibody, which had been physically absorbed on the wells of poly-styrene microplate and was used as a universal solid phase. The effect of several factors, such as the dilution ratios of the immunoreagents, the chemiluminescent substrate, the chemiluminescence reaction time and incu- bation condition were examined and optimized. The optimal dilutions of anti-progesterone antibody and HRP-progesterone conjugate were 1：10000 and 1：15000, respectively. The substrate Ⅱ was chosen and the luminescence was determined after 10 min incubation. The immunoreaction was incubated in water bath at 37 ℃ for 1 h. The assay was evaluated with sensitivity as low as 0.08 μg/L. The RSD was less than 15% for both intra-and inter-assay precision. The recoveries of three different spiked concentration samples were 101%, 101% and 94.4%, respectively. This proposed method has been successfully applied to the clinical evaluation of progesterone in 36 human sera. The results showed a good correlation with the commercially available radioimmunoassay kit with a correlative coefficient of 0. 9502.