摘要
探讨了对碘苯酚（PIP）增强的鲁米诺—过氧化氢—辣很过氧化物酶（Luminol-H_2O_2-HRP）体系的反应特性,通过正交试验并经方差分析,获得了增强发光酶免疫检测法测定辣根过氧化物酶（HRP）及其标记物的最佳实验条件（缓冲液的pH=8.2,Luminol的浓度5×10^(-4)mol/L,H_2O_2的浓度4×10^(-3)mol/L,PIP的浓度4×10^(-4)mol/L,乙二胺四乙酸的浓度0.25mg/ml,温度25℃.反应时间10min）。在上述最佳条件下检测HRP的浓度,其线性范围为:10^(-9)～10^(-12)g/L,Y=-0.05+1.07X（r=0.96,P<0.01）,最低检出限为 0.2pg（5 amol）。
The reaction characteristics of luminol- H2O2-horseradish peroxidase (HRP) system enhanced by paraiodophenol (PIP) were investigated. The optimal experimental conditions of enhanced chemiluminescence for free HRP and labeled HRP, which obtained by orthogonal design and analysis of variance, were found as follows: luminol buffer pH was 8. 2, the concentrations of luminol. H2O2,PIP and EDTA were 5×10-4 mol/L,4 ×10-3 mol/L ,4×10-4 mol/L and 0.25 mg/ml respectively, temperature was 25 C , and the reaction time was 10 min. Under these conditions, the working range of HRP was 10-9 to 10-12 g/L, and regression equation, Y=-0.05+1.07X(r=0.96,P<0.01) obtained. the minimum detection limit was 0.2×10-12g(5 amol).
出处
《同济医科大学学报》
CSCD
1998年第6期445-447,453,共4页
Acta Universitatis Medicinae Tongji
