Development of an ELISA for rapid detection of anti-type VII collagen autoantibodies in epidermolysis bullosa acquisita
Development of an ELISA for rapid detection of anti-type VII collagen autoantibodies in epidermolysis bullosa acquisita
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DOI：
10.1111/1523-1747.ep12285634
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发表时间：
1997-01-01
影响因子：
6.5
通讯作者：
Woodley, DT
中科院分区：
文献类型：
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作者：
Chen, M;Chan, LS;Woodley, DT
大疱性表皮松解症 (EBA) 是一种获得性水疱性皮肤病，其特征是存在针对 VII 型胶原蛋白的 IgG 自身抗体。 EBA 自身抗体可识别位于氨基末端非胶原 (NC1) 结构域内的四个主要免疫显性表位。在本研究中，我们开发了一种快速定量酶联免疫吸附测定 (ELISA)，使用真核表达的重组人 NC1 抗原检测针对 VII 型胶原完整 NC1 结构域的自身抗体活性。通过 ELISA，我们测试了 EBA 患者 (n = 24)、大疱性全身性血清红斑狼疮 (BSLE) (n = 3)、大疱性类天疱疮 (n = 16)、天疱疮 (n = 11) 和正常对照 (n = 12)。所有 EBA 和 BSLE 血清，包括间接免疫荧光阴性的四种血清，在 ELISA 中均表现出与固定化 NC1 的反应性。相比之下，来自健康对照受试者或患有无关水疱性皮肤病的患者的血清均未与 NC1 发生反应。通过免疫印迹分析，EBA 血清也与重组 NC1 发生反应，但灵敏度较低。因此，新开发的使用重组 NC1 的 ELISA 是一种灵敏、特异的测定方法，是快速筛选 EBA 和 BSLE 血清的有用工具。
Epidermolysis bullosa acquisita (EBA) is an acquired blistering skin disease characterized by the presence of IgG autoantibodies to type VII collagen. EBA autoantibodies recognize four major immunodominant epitopes localized within the amino-terminal, noncollagenous (NC1) domain, In this study, we developed a rapid, quantitative enzyme-linked immunosorbent assay (ELISA) to detect autoantibody activity against the complete NC1 domain of type VII collagen with the use of an eukaryotic-expressed, recombinant human NC1 antigen, With the ELISA, we tested serum from patients with EBA (n = 24), bullous systemic lupus erythematosus (BSLE) (n = 3), bullous pemphigoid (n = 16), pemphigus (n = 11), and normal controls (n = 12). All EBA and BSLE serum, including four sera that were negative by indirect immunofluorescence, demonstrated reactivity with immobilized NC1 in the ELISA, In contrast, none of the sera from healthy control subjects or patients with unrelated blistering skin diseases reacted with NC1. The EBA sera also reacted with recombinant NC1 by immunoblot analysis but with less sensitivity, Thus, the newly developed ELISA using recombinant NC1 is a sensitive, specific assay and a useful tool for rapidly screening EBA and BSLE serum.