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竞争性化学发光酶免疫分析法检测睾酮 |
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刘明泽 黄 岚 李振雪 徐 伟 张 旗 刘 杰
中国图书分类号 R392-33
摘 要 目的：建立高灵敏度的化学发光酶免疫分析法(CLEIA)检测睾酮。方法：包被抗体、标记抗原或包被抗原、标记抗体，用辣根过氧化物酶标记，鲁米诺增敏化学发光体系作为酶底物。结果：包被二抗、标记睾酮的方法灵敏度高、操作简便，灵敏度为0.063 ng/ml，睾酮浓度为1.0、5.0和10.0 ng/ml时，批内变异5.8%～7.4%(n=20)，批间变异为6.6%～8.7%(n=20)，回收率为98.2%～104.0%。与5α-二氢睾酮交叉反应为20.0%，与其它类固醇无明显交叉反应。此法与宝灵曼(BM)直接酶免疫分析试剂盒比较有较好的相关性，CLEIA=0.915 BM+0.598 ng/ml，n=128，r=0.971，P＜0.001。结论：此法灵敏度高，有较好的准确性和重复性，与商品试剂盒有较好的相关性，适合临床应用。
关键词 化学发光酶免疫分析法 睾酮 鲁米诺
Competitive chemiluminescent enzyme immunoassay for testosterone
LIU Ming-Ze, HUANG Lan, LI Zhen-Xue et al. Pilotech Institute of Biomedical Engeneering, People s Hospital, BMU, Beijing 100034
Abstract Objective: To establish a chemiluminescent enzyme immunoassay (CLEIA) for testosterone. Methods: Antibody was coated and antigen was labeled or antigen was coated and antibody was labeled. Horseradish peroxidase was used as a label and enhanced chemiluminescent system as substrate. Results: The method with coated second antibody and labeled testosterone had high sensitivity and simple measures. The sensitivity of the assay was 0.063 ng/ml. The testosterone was added to the steroid-free serum at concentrations of 1.0, 5.0 and 10.0 ng/ml, the within-run coefficients of variation were 5.8%～7.4%(n=20), and run-to-runs were 6.6%～8.7%(n=2)respectively. The recovery rates were from 98.2% to 104.0%. The assay was not cross-reacted significantly with other steroids except 5α-dihydrotestosterone with 20% cross reactivity. It correlated well with Boehringer Mannheim enzyme immunoassay kit (BM) for testosterone. CLEIA=0.915 BM+0.598 ng/ml, n=128, r=0.971. Conclusion: The chemiluminescent enzyme immunoassay has high sensitivity, good accuracy, repeatability and correlation with commercial kit. The method is helpful to clinical laboratory diagnosis.
Key words Chemiluminescent enzyme immunoassay Testosterone Luminol
血清中睾酮的测定有着十分重要的意义，在男性可以用于研究[1] [2] 下一页
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