【摘要】 目的:建立ELISA显色体系的优化方法,优化TMB/HPR显色系统的反应参数。方法:首先采用单因素试验,研究显色体系中的缓冲溶液pH、TMB浓度、H2O2浓度、反应时间和温度对显色的影响,在此基础上,再进行5因素4水平的正交试验。结果:TMB/HRP体系的最佳工作条件:缓冲液pH为5.0,TMB浓度为2.5mg/ml,H2O2浓度为0.03%(V/V),反应温度为30～50℃,反应时间为10min。结论:优化的TMB/HRP最佳反应条件,可以满足一般反应的要求,所建立的TMB/HRP体系的优化方法也适用于其它ELISA显色系统的优化。更多还原

【Abstract】 Objective: Establishing an optimization method for chromogenic substrate system of ELISA to optimize the reaction parameters of the TMB/HRP chromogenic substrate system.Method: Several single factor experiments were applied to analyze the effects of concentration of TMB and H2O2,pH,temperature and time on chromogenic reaction.Based on the single factor tests,the orthogonal experiment design method was use to further optimize reaction parameters.Result: The optimal reaction parameters were sodium acetate-citric acid buffer of pH 5.0,2.5mg/ml TMB,0.03%(V/V)H2O2,30-50℃ of reaction temperature and 10min of reaction.Conclusion: The optimal reaction parameters of TMB/HRP system were established,and the method was suitable for optimization of other chromogenic substrate systems of ELISA.更多还原