Usefulness of enzyme-linked immunosorbent assay using recombinant desmogleins 1 and 3 for serodiagnosis of pemphigus

Masayuki Amagai, A. Komai, T. Hashimoto, Y. Shirakata, K. Hashimoto, T. Yamada, Y. Kitajima, K. Ohya, H. Iwanami, T. Nishikawa

Research output: Contribution to journalArticle

306 Citations (Scopus)

Abstract

Pemphigus is an autoimmune blistering disease with two major subtypes, pemphigus vulgaris (PV) and pemphigus foltaceus (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 Dsg 1; if a serum is negative for Dsg3 and positive for Dsg1, it indicates a diagnosis of PF.

Original languageEnglish
Pages (from-to)351-357
Number of pages7
JournalBritish Journal of Dermatology
Volume140
Issue number2
DOIs
Publication statusPublished - 1999
Externally publishedYes

Fingerprint

Desmoglein 3
Desmoglein 1
Pemphigus
Serologic Tests
Enzyme-Linked Immunosorbent Assay
Serum
Collagen Diseases
Bullous Pemphigoid
Baculoviridae

Keywords

  • Desmoglein 1
  • Desmoglein 3
  • Diagnosis
  • Enzyme-linked immunosorbent assay
  • Pemphigus

ASJC Scopus subject areas

  • Dermatology

Cite this

Usefulness of enzyme-linked immunosorbent assay using recombinant desmogleins 1 and 3 for serodiagnosis of pemphigus. / Amagai, Masayuki; Komai, A.; Hashimoto, T.; Shirakata, Y.; Hashimoto, K.; Yamada, T.; Kitajima, Y.; Ohya, K.; Iwanami, H.; Nishikawa, T.

In: British Journal of Dermatology, Vol. 140, No. 2, 1999, p. 351-357.

Research output: Contribution to journalArticle

Amagai, M, Komai, A, Hashimoto, T, Shirakata, Y, Hashimoto, K, Yamada, T, Kitajima, Y, Ohya, K, Iwanami, H & Nishikawa, T 1999, 'Usefulness of enzyme-linked immunosorbent assay using recombinant desmogleins 1 and 3 for serodiagnosis of pemphigus', British Journal of Dermatology, vol. 140, no. 2, pp. 351-357. https://doi.org/10.1046/j.1365-2133.1999.02752.x
Amagai, Masayuki ; Komai, A. ; Hashimoto, T. ; Shirakata, Y. ; Hashimoto, K. ; Yamada, T. ; Kitajima, Y. ; Ohya, K. ; Iwanami, H. ; Nishikawa, T. / Usefulness of enzyme-linked immunosorbent assay using recombinant desmogleins 1 and 3 for serodiagnosis of pemphigus. In: British Journal of Dermatology. 1999 ; Vol. 140, No. 2. pp. 351-357.
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abstract = "Pemphigus is an autoimmune blistering disease with two major subtypes, pemphigus vulgaris (PV) and pemphigus foltaceus (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 Dsg 1; if a serum is negative for Dsg3 and positive for Dsg1, it indicates a diagnosis of PF.",
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AU - Komai, A.

AU - Hashimoto, T.

AU - Shirakata, Y.

AU - Hashimoto, K.

AU - Yamada, T.

AU - Kitajima, Y.

AU - Ohya, K.

AU - Iwanami, H.

AU - Nishikawa, T.

PY - 1999

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AB - Pemphigus is an autoimmune blistering disease with two major subtypes, pemphigus vulgaris (PV) and pemphigus foltaceus (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 Dsg 1; if a serum is negative for Dsg3 and positive for Dsg1, it indicates a diagnosis of PF.

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KW - Enzyme-linked immunosorbent assay

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