Jones J V, Mansour M, James H, Sadi D, Carr R I
Department of Medicine, Dalhousie University, Halifax, Nova Scotia, Canada.
J Immunol Methods. 1989 Mar 10;118(1):79-84. doi: 10.1016/0022-1759(89)90056-2.
We have recently described a substrate amplification system, based on the method of Self, which increases the sensitivity of alkaline phosphatase (AP)-dependent enzyme-linked immunosorbent assays (ELISA) by a factor of 30-50. This increase is achieved by having AP, the primary enzyme, produce an activator for a secondary enzyme-substrate system, within which marked amplification occurs. We have now demonstrated that this amplification method can be applied to the measurement of human antibodies to DNA. The sensitivity is greater by a factor of 10 than the conventional method, which uses p-nitrophenyl phosphate (p-NPP) as substrate. On replicate assays the method is reproducible, with a coefficient of variation of less than 0.1. This great increase in sensitivity should be of value in conserving specimens of serum and in screening monoclonal antibodies.
我们最近描述了一种基于塞尔夫方法的底物扩增系统，该系统可将碱性磷酸酶（AP）依赖性酶联免疫吸附测定（ELISA）的灵敏度提高30至50倍。这种提高是通过让主要酶AP产生一种用于二级酶-底物系统的激活剂来实现的，在该系统中会发生显著的扩增。我们现在已经证明，这种扩增方法可用于测量人抗DNA抗体。其灵敏度比使用对硝基苯磷酸酯（p-NPP）作为底物的传统方法高10倍。在重复测定中，该方法具有可重复性，变异系数小于0.1。灵敏度的大幅提高在保存血清标本和筛选单克隆抗体方面应具有价值。
