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用于I型过敏症中组分分辨诊断(CRD)的微阵列技术。一种处于研究工具和常规诊断之间边界的创新技术

[Microarray technique for component resolved diagnosis (CRD) in type-I allergies. An innovative technology at the border between research tool and routine diagnostics].

作者信息

Klimek L, Vetter D, von Bernus L, Thorn C

机构信息

Zentrum für Rhinologie und Allergologie Wiesbaden, An den Quellen 10, 65183, Wiesbaden, Deutschland.

出版信息

HNO. 2011 Oct;59(10):988-93. doi: 10.1007/s00106-010-2224-5.

Abstract

Approximately 25% of the population in industrialized countries suffer from IgE-associated Type-1 allergies. Multiple allergens can be tested simultaneously in one assay by using the protein microarray. Moreover, it is possible to measure more than one analytical parameter (e.g. allergen specific IgEs and IgGs) in one assay by combining different fluorescent markers with specific secondary antibodies. The different allergen components that are of interest are immobilized on a planar surface. By adding the patient's serum (a smaller amount of serum is needed compared to an immunoassay) the inherent IgE antibodies are captured by the corresponding allergens. Secondary fluorescing anti-IgE antibodies are added subsequently, thus the intensity of each spot on the microarray can be measured by using a biochipscanner. The detected signal is then transformed into quantitative data, which allows the classification of the patient's serum IgE level for the tested allergens. There are different approaches to reduce the complexity of the original extracts used for the production of the solid microarray phase to a smaller number of relevant pathogenic molecules. The component-resolved diagnosis still needs to be clinically validated, but initial studies show positive results concerning the sensitivity and specificity of the protein microarray. Protein microarrays are promising tools for screening diagnoses in allergic diseases as well as for the improvement of allergen-specific immunotherapy.

摘要

在工业化国家,约25%的人口患有与IgE相关的1型过敏症。通过使用蛋白质微阵列,可以在一次检测中同时检测多种过敏原。此外,通过将不同的荧光标记物与特异性二抗结合,在一次检测中可以测量多个分析参数(如过敏原特异性IgE和IgG)。将感兴趣的不同过敏原成分固定在平面表面上。加入患者血清(与免疫测定相比,所需血清量较少)后,固有IgE抗体被相应的过敏原捕获。随后加入荧光二抗抗IgE抗体,因此可以使用生物芯片扫描仪测量微阵列上每个斑点的强度。然后将检测到的信号转化为定量数据,从而对患者血清中针对所检测过敏原的IgE水平进行分类。有不同的方法可将用于制备固相微阵列相的原始提取物的复杂性降低到较少数量的相关致病分子。组分分辨诊断仍需临床验证,但初步研究显示了蛋白质微阵列在敏感性和特异性方面的积极结果。蛋白质微阵列是用于过敏性疾病筛查诊断以及改善过敏原特异性免疫疗法的有前景的工具。

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