Usefulness of enzyme‐linked immunosorbent assay using recombinant desmogleins 1 and 3 for serodiagnosis of pemphigus
Usefulness of enzyme‐linked immunosorbent assay using recombinant desmogleins 1 and 3 for serodiagnosis of pemphigus
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DOI：
10.1046/j.1365-2133.1999.02752.x
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发表时间：
1999-02
影响因子：
10.3
通讯作者：
M. Amagai;A. Komai;T. Hashimoto;Y. Shirakata;K. Hashimoto;T. Yamada;Y. Kitajima;K. Ohya;H. Iwanami;T. Nishikawa
中科院分区：
文献类型：
--
作者：
M. Amagai;A. Komai;T. Hashimoto;Y. Shirakata;K. Hashimoto;T. Yamada;Y. Kitajima;K. Ohya;H. Iwanami;T. Nishikawa
天疱疮是一种自身免疫性水疱性疾病，有两种主要亚型，寻常型天疱疮（PV）和落叶型天疱疮（PF）。天疱疮患者存在循环抗桥粒芯糖蛋白（Dsg）1和/或抗Dsg 3 IgG自身抗体。我们之前已经开发了使用杆状病毒表达的重组Dsg 1和Dsg 3作为天疱疮诊断工具的酶联免疫吸附试验（ELISA）。本研究的目的是评价这些ELISA在临床上大量血清样本的实际应用。我们使用了81份PV血清、48份PF血清、114份大疱性类天疱疮（BP）血清、124份胶原病血清、9份其他非天疱疮大疱性疾病血清和179份正常对照血清。通过受试者工作特征图确定临界值。48份PF血清中有47份（97.9%）在Dsg 1 ELISA中呈阳性，81份PV血清中有79份（97.5%）在Dsg 3 ELISA中呈阳性，而179份正常血清中分别仅有2份（1.1%）和4份（2.2%）在Dsg 1和Dsg 3 ELISA中呈阳性。然而，BP和胶原蛋白疾病的一些疾病对照血清超过临界值。灰色区域的引入有助于减少这些假阳性血清的数量。此外，在研究的三名患者中，各自的Dsg 1和Dsg 3 ELISA评分显示出沿着时间进程与疾病活动平行波动。我们的结论是，Dsg 1和Dsg 3 ELISA为PV和PF患者的诊断提供了一种简单，敏感和高度特异性的检测方法，这些ELISA可能是一个有价值的工具，以监测疾病的活动。我们还提出了基于ELISA反应性的天疱疮诊断标准：如果血清对Dsg 3呈阳性，则表明诊断为PV，而不考虑对Dsg 1的反应性;如果血清对Dsg 3呈阴性而对Dsg 1呈阳性，则表明诊断为PF。
Pemphigus is an autoimmune blistering disease with two major subtypes, pemphigus vulgaris (PV) and pemphigus foliaceus (PF). Patients with pemphigus have circulating antidesmoglein (Dsg)1 and/or anti‐Dsg3 IgG autoantibodies. We have previously developed enzyme‐linked immunosorbent assays (ELISAs) using recombinant Dsg1 and Dsg3 expressed by baculovirus as a diagnostic tool for pemphigus. The purpose of this study was to evaluate the practical application of these ELISAs for clinical use with a large number of serum samples. We used 81 PV sera, 48 PF sera, 114 bullous pemphigoid (BP) sera, 124 collagen disease sera, nine sera of other non‐pemphigus bullous diseases and 179 normal control sera. A cut‐off value was determined by receiver‐operating‐characteristic plots. Forty‐seven of 48 PF sera (97.9%) were positive in the Dsg1 ELISA and 79 of 81 PV sera (97.5%) were positive in the Dsg3 ELISA, while only two (1.1%) and four (2.2%) of 179 normal sera were positive in Dsg1 and Dsg3 ELISAs, respectively. However, some disease control sera of BP and collagen diseases exceeded the cut‐off value. Introduction of a grey zone helped to decrease the number of these false‐positive sera. Furthermore, in three patients studied, the respective Dsg1 and Dsg3 ELISA scores showed parallel fluctuation with the disease activity along the time course. We conclude that Dsg1 and Dsg3 ELISAs provide a simple, sensitive and highly specific assay for the diagnosis of patients with PV and PF and that these ELISAs may be a valuable tool to monitor the disease activity. We also propose diagnostic criteria for pemphigus based on ELISA reactivity: if a serum is positive against Dsg3 it indicates a diagnosis of PV, regardless of reactivity against Dsg1; if a serum is negative for Dsg3 and positive for Dsg1, it indicates a diagnosis of PF.