Biochip technology for the serological diagnosis of bullous pemphigoid.

ISRN Dermatology Pub Date : 2012-01-01 Epub Date: 2012-12-17 DOI:10.5402/2012/237802
Haik Zarian, Andrea Saponeri, Anna Michelotto, Edoardo Zattra, Anna Belloni-Fortina, Mauro Alaibac
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引用次数: 34

Abstract

Bullous pemphigoid is an autoimmune blistering skin disease characterized by the presence of circulating autoantibodies which recognize specific proteins of the epidermis and dermoepidermal junction. Diagnosis is based on clinical criteria and laboratory investigations, notably histology, direct and indirect immunofluorescence, and ELISA. This study describes a new immunofluorescence assay for parallel determination of anti-BP180 and anti-BP230 based on recombinant antigenic substrates. The aim of the study was to detect BP180 and BP230 autoantibodies by BIOCHIP technology using both a specially designed recombinant BP180-NC16A protein and cells expressing the BP230-gc antigen fragment. 18 patients with bullous pemphigoid were included in the study. Autoantibodies to BP180 were detected by the BIOCHIP technique in 83.33% of patients with clinical, serological, and immunohistological confirmed bullous pemphigoid while autoantibodies against BP230-gC were detected only in 39% of patients. The detection of anti-BP180-NC16A and anti-BP230-gC by a new biochip-based immunoassay is a suitable alternative to indirect immunofluorescence and ELISA. This method has the advantage of easily discriminating the different autoantibody specificities. The BIOCHIP method is faster, cheaper, and easy to use when compared with the ELISA approach. For this reason, the new method could be used as an initial screening test to identify patients with bullous pemphigoid, and doubtful results could then be confirmed by ELISA.

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生物芯片技术用于大疱性类天疱疮的血清学诊断。
大疱性类天疱疮是一种自身免疫性起泡性皮肤病,其特征是存在循环自身抗体,该抗体可识别表皮和真皮表皮交界处的特定蛋白质。诊断是基于临床标准和实验室调查,特别是组织学,直接和间接免疫荧光,和ELISA。本研究描述了一种新的基于重组抗原底物的平行测定抗bp180和抗bp230的免疫荧光法。本研究的目的是利用BIOCHIP技术,利用专门设计的重组BP180- nc16a蛋白和表达BP230-gc抗原片段的细胞,检测BP180和BP230自身抗体。18例大疱性类天疱疮患者纳入研究。83.33%的临床、血清学和免疫组织学证实的大疱性类天疱疮患者通过BIOCHIP技术检测到BP180自身抗体,而仅39%的患者检测到BP230-gC自身抗体。基于生物芯片的新型免疫分析法检测抗bp180 - nc16a和抗bp230 - gc是间接免疫荧光和ELISA的一种合适的替代方法。该方法具有容易区分不同的自身抗体特异性的优点。与ELISA方法相比,BIOCHIP方法更快、更便宜、更容易使用。因此,新方法可作为鉴别大疱性类天疱疮患者的初步筛选试验,可疑的结果可通过ELISA进行确认。
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