竞争性化学发光酶免疫分析法检测睾酮

Competitive chemiluminescent enzyme immunoassay for testosterone

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０６３ｎｇ／ｍｌ，睾酮浓度为１０、５０和１００ｎｇ／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９１５ＢＭ＋０５９８ｎｇ／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。

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 steroidfree serum at concentrations of 1.0, 5.0 and 10.0 ng/ml, the withinrun coefficients of variation were 5. 8%7.4%( n =20), and runtoruns' were 6.6%8.7%( n =2)respectively. The recovery rates were from 98.2% to 104.0%. The assay was not crossreacted significantly with other steroids except 5dihydrotestosterone 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.