Institute of Experimental Immunology, EUROIMMUN AG, Luebeck, Germany.
J Am Acad Dermatol. 2013 Mar;68(3):e89-95. doi: 10.1016/j.jaad.2011.12.032. Epub 2012 Feb 16.
Epidermolysis bullosa acquisita (EBA) is a severe autoimmune subepidermal blistering disease characterized by autoantibodies against the N-terminal collagenous domain (NC1) of type VII collagen (Col VII).
Development of reliable assays for the detection of anti-Col VII-NC1 antibodies.
NC1 was expressed in human HEK293 cells and used as target antigen in an enzyme-linked immunosorbent assay (ELISA) and in an immunofluorescence assay (IFA). These two assays were probed in a large cohort of patients with EBA (n = 73), bullous pemphigoid (BP, n = 72), anti-p200 pemphigoid (n = 24), anti-laminin 332 mucous membrane pemphigoid (MMP, n = 15), pemphigus vulgaris (PV, n = 24), and healthy control subjects (n = 254).
The cut-off for the ELISA was optimized for accuracy by receiver-operating characteristics (area under the curve [AUC] = 0.9952). IgG reactivity against NC1 was detected in 69 of 73 EBA (94.5%) and 5 control sera (2 healthy controls and 3 BP patients), resulting in a specificity of 98.7%. The IFA showed a sensitivity of 91.8% and specificity of 99.8%. Reproducibility of the ELISA was demonstrated by an intra-class correlation coefficient of 0.97. IgG subclass analyses by ELISA revealed IgG1, IgG2, IgG3, and IgG4 anti-NC1 reactivity in 83.6%, 85.3%, 37.7%, and 83.6% of EBA sera, respectively.
The novel assays were not evaluated prospectively and their use in monitoring serum levels during the disease course was not tested.
The two assays are highly specific and sensitive to diagnose EBA. Their diagnostic competence was demonstrated in a large cohort of well-characterized EBA sera.
获得性大疱性表皮松解症（EBA）是一种严重的自身免疫性表皮下水疱性疾病，其特征是存在针对 VII 型胶原（Col VII）N 端胶原域（NC1）的自身抗体。
开发用于检测抗 Col VII-NC1 抗体的可靠检测方法。
在人 HEK293 细胞中表达 NC1，并将其作为酶联免疫吸附测定（ELISA）和免疫荧光测定（IFA）的靶抗原。在 73 例 EBA 患者、72 例大疱性类天疱疮（BP）患者、24 例抗 p200 类天疱疮患者、15 例抗层粘连蛋白 332 黏膜性大疱性皮病（MMP）患者、24 例寻常性天疱疮患者和 254 例健康对照者的大样本队列中，对这两种检测方法进行了检测。
通过接收者操作特征（曲线下面积[AUC] = 0.9952）对 ELISA 的截止值进行了优化，以确保准确性。在 73 例 EBA 患者中有 69 例（94.5%）和 5 例对照血清（2 例健康对照和 3 例 BP 患者）检测到针对 NC1 的 IgG 反应，特异性为 98.7%。IFA 的敏感性为 91.8%，特异性为 99.8%。通过内部类间相关系数 0.97 证明了 ELISA 的可重复性。通过 ELISA 进行 IgG 亚类分析，分别在 83.6%、85.3%、37.7%和 83.6%的 EBA 血清中检测到 IgG1、IgG2、IgG3 和 IgG4 抗 NC1 反应性。
这两项新检测方法均未进行前瞻性评估，也未对其在疾病过程中监测血清水平的应用进行测试。
这两种检测方法对诊断 EBA 具有高度的特异性和敏感性。它们在一组特征明确的 EBA 血清中的诊断能力得到了证实。
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