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过敏原微阵列:使用重组过敏原的微阵列与传统诊断方法检测过敏原特异性血清免疫球蛋白E的比较。

Allergen microarray: comparison of microarray using recombinant allergens with conventional diagnostic methods to detect allergen-specific serum immunoglobulin E.

作者信息

Jahn-Schmid B, Harwanegg C, Hiller R, Bohle B, Ebner C, Scheiner O, Mueller M W

机构信息

Institute of Pathophysiology, University of Vienna, Vienna, Austria.

出版信息

Clin Exp Allergy. 2003 Oct;33(10):1443-9. doi: 10.1046/j.1365-2222.2003.01784.x.

Abstract

BACKGROUND

The availability of recombinant allergens and recent advances in biochip technology led to the development of a novel test system for the detection of allergen-specific IgE.

OBJECTIVE

To test the performance of this allergen microarray in a serological analytical study.

METHODS

Standard allergens contained in grass pollen (Phl p 1, Phl p 2, Phl p 5 and Phl p 6) and tree pollen (Bet v 1 and Bet v 2) were used as a model system. The detection of allergen-specific serum IgE using microarrays was compared with standard test systems: CAP/RAST and an in-house ELISA. In order to test the analytical sensitivity of the assays, geometric dilutions of a serum pool containing high levels of pollen-specific IgE from allergic individuals were tested in each system. To assess the analytical specificity, the sera of 51 patients with presumptive allergic symptoms were collected before diagnosis. Thereafter, the results for grass/tree-pollen-specific IgE were compared.

RESULTS

The microarray has a good dynamic range similar to the CAP/RAST system. Microarray and ELISA showed comparable analytical sensitivity exceeding the CAP/RAST system. With respect to the analytical specificity, no significant cross-reactivity of the allergens was observed. For two of the allergens tested, weak positive signals were detected in the microarray test system, whereas they were not detectable by CAP/RAST.

CONCLUSION

A good correlation of presently used methods to detect serum IgE and the novel microarray test system was observed. As a next step, a careful validation of this method for a multitude of allergens and a thorough clinical evaluation has to be provided. Microarray testing of allergen-specific IgE can be presumed to be the method of choice for a prospective component-resolved diagnosis of Type I allergy, and the basis for the design and monitoring of a patient-tailored specific immunotherapy in the future.

摘要

背景

重组变应原的可得性以及生物芯片技术的最新进展促使开发出一种用于检测变应原特异性IgE的新型检测系统。

目的

在一项血清学分析研究中测试这种变应原微阵列的性能。

方法

以草花粉(Phl p 1、Phl p 2、Phl p 5和Phl p 6)和树花粉(Bet v 1和Bet v 2)中含有的标准变应原为模型系统。将使用微阵列检测变应原特异性血清IgE与标准检测系统进行比较:CAP/RAST和一种内部ELISA。为了测试这些检测方法的分析灵敏度,在每个系统中对含有来自过敏个体的高水平花粉特异性IgE的血清池进行几何稀释检测。为了评估分析特异性,在诊断前收集了51例疑似过敏症状患者的血清。此后,比较草/树花粉特异性IgE的检测结果。

结果

微阵列具有与CAP/RAST系统相似的良好动态范围。微阵列和ELISA显示出相当的分析灵敏度,超过了CAP/RAST系统。在分析特异性方面,未观察到变应原的显著交叉反应。对于所检测的两种变应原,在微阵列检测系统中检测到弱阳性信号,而CAP/RAST未检测到。

结论

观察到目前用于检测血清IgE的方法与新型微阵列检测系统具有良好的相关性。下一步,必须对该方法针对多种变应原进行仔细验证并进行全面的临床评估。变应原特异性IgE的微阵列检测可被认为是I型过敏前瞻性组分分辨诊断的首选方法,也是未来设计和监测患者个体化特异性免疫疗法的基础。

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