Little S A, Warner J O
University of Southampton, England.
J Allergy Clin Immunol. 1996 Jul;98(1):55-63. doi: 10.1016/s0091-6749(96)70226-9.
In cystic fibrosis (CF) serologic tests assume increased significance for discriminating the symptoms of allergic bronchopulmonary aspergillosis (ABPA) from those of bacterial infections. Elevated specific IgE is a sensitive indicator of ABPA, but the high prevalence of patients with CF who have simple allergy to Aspergillus fumigatus means that it lacks specificity.
The aim of this study was to develop an assay to detect IgE to a single allergenic protein of A. fumigatus to increase the discrimination between ABPA and uncomplicated A. fumigatus allergy.
to identify a more selective allergen, serum samples from nine patients with ABPA and five patients with allergy were used to screen A. fumigatus proteins for allergenic activity by crossed radioimmunoelectrophoresis. Molecular weights were determined by electrophoresis and immunoblotting, and carbohydrate content was analyzed by concanavalin A chromatography and endoglycosidase digestion.
The allergen identified was found to be a glycoprotein with a molecular weight of 66 kd, referred to as gp66. An ELISA was developed to measure specific IgE to gp66, and the efficacy was compared with IgE detection by using the unfractionated A. fumigatus extract on serum samples from 14 patients with CF and ABPA, 42 patients with CF and allergy but not ABPA, and 23 patients with CF without ABPA or allergy.
Use of gp66 increased the specificity of IgE analysis from 58.5% to 78.5%, whereas the sensitivity of 92.9% was maintained.
在囊性纤维化(CF)中,血清学检测对于区分变应性支气管肺曲霉病(ABPA)和细菌感染的症状具有越来越重要的意义。特异性IgE升高是ABPA的敏感指标,但CF患者中对烟曲霉单纯过敏的高患病率意味着它缺乏特异性。
本研究的目的是开发一种检测针对烟曲霉单一变应原蛋白的IgE的检测方法,以提高ABPA与单纯烟曲霉过敏之间的鉴别能力。
为了鉴定一种更具选择性的变应原,使用来自9例ABPA患者和5例过敏患者的血清样本,通过交叉放射免疫电泳筛选烟曲霉蛋白的变应原活性。通过电泳和免疫印迹测定分子量,并通过伴刀豆球蛋白A色谱法和内切糖苷酶消化分析碳水化合物含量。
鉴定出的变应原是一种分子量为66kd的糖蛋白,称为gp66。开发了一种ELISA来测量针对gp66的特异性IgE,并将其与使用未分级的烟曲霉提取物检测14例CF合并ABPA患者、42例CF合并过敏但无ABPA患者以及23例无ABPA或过敏的CF患者血清样本中的IgE的功效进行比较。
使用gp66可将IgE分析的特异性从58.5%提高到78.5%,同时保持92.9%的敏感性。