间接酶联免疫吸附试验显色系统主要影响因素的探讨

The approach on the main influencing facters of 3, 3’, 5, 5’-tetramethylbenzidine substrate reagent solution system for an indirect ELISA

关键词:  酶联免疫吸附试验  过氧化脲  四甲基联苯胺  辣根过氧化物酶

Key words:  Enzyme linked immunosorbent assay  ELISA  Perhydrol-Urea  3,3,5,5-tetramethylbenzidine  TMB  HRP

摘要:

酶联免疫吸附试验（ELISA）易受多种因素影响，其中包括显色系统。本文就ELISA常用的四甲基联苯胺（TMB）显色系统的成份、含量及反应条件对ELISA显色状态的影响进行了探讨，制订了一种通用的调整包括pH值、底物成份及含量、反应时间等在内的显色系统参量的方法，应用该方法为确定适用于任一特定ELISA的显色系统以及建立稳定的ELISA体系奠定了基础。 关键词： 酶联免疫吸附试验；过氧化脲； 四甲基联苯胺；辣根过氧化物酶

Abstract:

The results of the different enzyme linked immunosorbent assay (ELISA) can be easily affected by many factors which includes chromogenic substrate system. Our research discussed on the influences of chromogenic condition by ingredients, contents, and reaction conditions of the commonly used TMB substrate reagent solution system, and finally made a general adjustment to the parameters of the chromogenic substrate system, including the pH, substrate ingredients, contents and reaction time. And by using this way we established a foundation to define a chromogenic substrate system for each different ELISA and to establish a stable ELISA system.