青霉素酶标记抗睾丸酮酶联免疫吸附分析法的研究

Studies of Enzyme-linked Immunoassay for Testosterone Using Penicillinase as Label

作者采用青霉素酶作标记酶，硝酸纤维素膜作免疫吸附载体，青霉素Ｖ－淀粉－碘试剂作底物，建立了一种测定睾九酮的酶联免疫吸附分析法。该法最低检出量为０．５ｎｇ／ｍｌ（２５ｐｇ／孔），其睾丸酮含量在１０￣（－２）～１０￣（－７）ｍｇ／ｍｌ范围内与吸光度值呈线性关系（ｒ＝０．９９６９），日内、日间变异系数（ＣＶ）分别小于３．３％（ｎ＝８）、５．９％（ｎ＝８），与放射免疫分析法检测之间有良好的相关性（ｒ＝０．９３７）

An enzyme-inked immunoassay(EIA)for testosterone was developed based on penicillinaseas the marker enzyme. The assay proposed requires aconjugate containing penicillinase as the enzymaticcomponent,penicillin Vas substrate and an iodine-starch reagent as the chromogenic component.Thebinding of conjugate molecules results in the transitionof the iodic complex from dark blue to colourless。△A(the difference between the blank and thetestosterone) vs log C was linear over the range 10￣ (-7)-10￣(-2)mg/ml(r=0.9969),with a limit of detection 25pg per disc。 The CVs of day to day and within day forplasma testosterone were<3。3%and5.9%, respec-tively。 There was a good correlation between thevalues obtained by EM and radioimmunoassay(r=0.937.)