Development of NC1 and NC2 domains of Type VII collagen ELISA for the diagnosis and analysis of the time course of epidermolysis bullosa acquisita patients

Marwah Adly Saleh, Ken Ishii, Yool Ja Kim, Akihiro Murakami, Norito Ishii, Takashi Hashimoto, Enno Schmidt, Detlef Zillikens, Yuji Shirakata, Koji Hashimoto, Yasuo Kitajima, Masayuki Amagai

研究成果: Article査読

81 被引用数 (Scopus)

抄録

Background: Epidermolysis bullosa acquisita (EBA) is an acquired autoimmune mechanobullous disease. EBA patients possess autoantibodies against type VII collagen which is composed of a collagenous domain flanked by non-collagenous NC1 and NC2 domains. It was reported that major epitopes reside within the NC1 domain and minor epitopes reside within NC2 domain. Objective: The aim of this study is to develop a sensitive and specific ELISA to facilitate the diagnosis of EBA. Methods: We developed ELISAs using recombinant NC1 domain produced by mammalian expression system and recombinant NC2 domain produced by mammalian or bacterial expression system to characterize autoantibodies in EBA. Next, we developed an ELISA using a combination of the NC1 (mammalian expression) and NC2 (bacterial expression). We tested the ELISAs with 49 EBA sera, 55 normal control sera, 20 pemphigus vulgaris and 20 bullous pemphigoid sera. Results: When we evaluated the 49 EBA sera using the NC1 and NC2 ELISAs, 38 (77.5%) reacted with NC1 domain only, 7 sera (14.2%) reacted with both NC1 and NC2 domains, and one serum (2%) reacted with NC2 domain only. Therefore, to increase the sensitivity of the assay, we developed an ELISA coated with a mixture of recombinant NC1 and NC2 domains, resulting in 93.8% sensitivity and 98.1% specificity. By analyzing the time course of two EBA patients, ELISA scores fluctuated in parallel with their disease activity. Conclusion: We conclude that the NC1. +. NC2 ELISA can be a practical assay for the diagnosis and follow up of the antibody titers of EBA patients.

本文言語English
ページ(範囲)169-175
ページ数7
ジャーナルJournal of Dermatological Science
62
3
DOI
出版ステータスPublished - 2011 6月

ASJC Scopus subject areas

  • 生化学
  • 分子生物学
  • 皮膚病学

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