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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-H_2O_2-HRP system enhanced by iodophenol (PIP) were investigated. By orthogonal test and variance analysis, The optimal conditions for the determination of horseradish peroxidase (HRP) and its markers by immunoassay (buffer pH = 8.2, Luminol concentration 5 × 10 -4 mol / L, H 2 O 2 concentration 4 × 10 ~ (-3) mol / L, PIP concentration 4 × 10 -4 mol / L, ethylenediaminetetraacetic acid concentration 0.25 mg / ml, temperature 25 ℃, reaction time 10 min). The linear range of HRP was 10 ~ (-9) ~ 10 ~ (-12) g / L and Y = -0.05 + 1.07X (r = 0.96, P <0.01) under the above optimal conditions. The minimum detection limit is 0.2pg (5 amol).