Falagiani P, Mistrello G, Rapisarda G, Festa A, Cislaghi C, Zanoni D
Research Department, Laboratorio Farmaceutico Lofarma S.R.l., MIlan, Italy.
J Immunol Methods. 1994 Aug 1;173(2):181-90. doi: 10.1016/0022-1759(94)90298-4.
The potency of allergenic extracts can be determined in vitro by RAST inhibition, and this has become the preferred method for the standardization of allergens. A disadvantage of this technique is the impossibility of obtaining data about allergens bound to the solid phase, i.e., the counterpart of the inhibiting extract. The REAST (reverse enzyme allergosorbent test) is based on the capture of IgE by a specific antibody bound to microtiter wells, the reaction of captured IgE with biotinylated allergen and the development of a colour reaction by subsequent addition of streptavidin-peroxidase and chromogenic substrate. The addition of an allergen extract in a dose-response fashion competes with the biotinylated allergen and inhibits the test. In the present study REAST inhibition has been evaluated with Dermatophagoides pteronyssinus, Parietaria judaica and mixed grass pollen extracts. The correlation of REAST inhibition with RAST inhibition and both intra-assay and inter-assay reproducibility have been evaluated. REAST inhibition is a potentially valuable new tool for the standardization of allergenic extracts.
变应原提取物的效价可通过放射性变应原吸附抑制试验（RAST抑制试验）在体外进行测定，这已成为变应原标准化的首选方法。该技术的一个缺点是无法获得与固相结合的变应原的数据，即抑制提取物的对应物的数据。反向酶变应原吸附试验（REAST）基于以下原理：通过与微量滴定板孔结合的特异性抗体捕获IgE，捕获的IgE与生物素化变应原反应，随后加入链霉亲和素-过氧化物酶和显色底物产生显色反应。以剂量反应方式加入变应原提取物会与生物素化变应原竞争并抑制试验。在本研究中，已使用屋尘螨、墙草和混合草花粉提取物对REAST抑制试验进行了评估。还评估了REAST抑制试验与RAST抑制试验的相关性以及试验内和试验间的重现性。REAST抑制试验是变应原提取物标准化的一种潜在有价值的新工具。
