Der f2重组蛋白用于变应性鼻炎患者血清ELISA检测

An Indirect ELISA Assay Using Recombinant Der f2 as Capture for the Analysis of Der f2-specific IgE in Perennial Allergic Rhinitis Patients

目的:以重组表达的野生型粉尘螨变应原蛋白质第2组(Derf2)分作为固相包被抗原,建立间接酶联免疫吸附试验(ELISA)法,检测粉尘螨致敏的持续性变应性鼻炎患者血清中的特异性IgE浓度。方法:经过皮肤点刺试验证明的24例粉尘螨反应阳性血清和2例阴性血清作为IgE血清样本。以粉尘螨变应原蛋白质第2组分固相包被酶标板,辣根过氧化物酶(HRP)标记的抗人IgE单抗作为第2抗体组成测试系统分析特异性IgE。结果:间接ELISA系统检测灵敏度为0.73U.mL-1。在4.0U.mL-1、12.0U.mL-1、22.0U.mL-1浓度,批内重复性和批间重复性分别为9.5%,7%,6.1%和12.8%,9.1%,7.2%。24例皮试阳性患者血清经ELISA分析,有22例阳性。结论:以粉尘螨变应原蛋白质第2组分作为固相包被抗原的间接ELISA法可用来对患者血清sIgE作定量或定性分析,有助于持续性变应性鼻炎的实验诊断。

Objective：To develop an indirect ELISA assay which uses recombinant Der f2 as capture for the analysis of Der f2-specific IgE concentration in sera of perennial allergic rhinitis patients. Methods：A panel of 24 sera of Der f-allergic patients and 2 control sera confirmed by skin prick test were used for the assays. Der f2-specific IgE was assayed using plates coated with the purified recombinant Der f2 allergen. IgE binding was detected with horseradish peroxidase-conjugated anti-human IgE monoclonal antibody as the secondary antibody. Results：The sensitivity of the indirect ELISA system was 0.73 U·mL-1,and the precision levels were 9.5%,7%,6.1% for intraassay and 12.8%,9.1%,7.2% for interassay at the concentration of 4 U·mL-1,12 U·mL-1 and 22 U·mL-1,respectively. Der f2-sIgEs were positive in 22/24 skin prick test positive patient sera.Conclusions：The established indirect ELISA test proved to be a valuable tool for determination of Der f2-sIgE and for evaluation of sIgE during clinical course of allergic rhinitis.