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Sensitive and specific assays for routine serological diagnosis of epidermolysis bullosa acquisita.
J Am Acad Dermatol. 2013 Mar; 68(3):e89-95.JA

Abstract

BACKGROUND

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).

OBJECTIVE

Development of reliable assays for the detection of anti-Col VII-NC1 antibodies.

METHODS

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).

RESULTS

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.

LIMITATIONS

The novel assays were not evaluated prospectively and their use in monitoring serum levels during the disease course was not tested.

CONCLUSION

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.

Authors+Show Affiliations

Institute of Experimental Immunology, EUROIMMUN AG, Luebeck, Germany.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22341608

Citation

Komorowski, Lars, et al. "Sensitive and Specific Assays for Routine Serological Diagnosis of Epidermolysis Bullosa Acquisita." Journal of the American Academy of Dermatology, vol. 68, no. 3, 2013, pp. e89-95.
Komorowski L, Müller R, Vorobyev A, et al. Sensitive and specific assays for routine serological diagnosis of epidermolysis bullosa acquisita. J Am Acad Dermatol. 2013;68(3):e89-95.
Komorowski, L., Müller, R., Vorobyev, A., Probst, C., Recke, A., Jonkman, M. F., Hashimoto, T., Kim, S. C., Groves, R., Ludwig, R. J., Zillikens, D., Stöcker, W., & Schmidt, E. (2013). Sensitive and specific assays for routine serological diagnosis of epidermolysis bullosa acquisita. Journal of the American Academy of Dermatology, 68(3), e89-95. https://doi.org/10.1016/j.jaad.2011.12.032
Komorowski L, et al. Sensitive and Specific Assays for Routine Serological Diagnosis of Epidermolysis Bullosa Acquisita. J Am Acad Dermatol. 2013;68(3):e89-95. PubMed PMID: 22341608.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Sensitive and specific assays for routine serological diagnosis of epidermolysis bullosa acquisita. AU - Komorowski,Lars, AU - Müller,Ralf, AU - Vorobyev,Artem, AU - Probst,Christian, AU - Recke,Andreas, AU - Jonkman,Marcel F, AU - Hashimoto,Takashi, AU - Kim,Soo-Chan, AU - Groves,Richard, AU - Ludwig,Ralf J, AU - Zillikens,Detlef, AU - Stöcker,Winfried, AU - Schmidt,Enno, Y1 - 2012/02/16/ PY - 2011/09/02/received PY - 2011/12/16/revised PY - 2011/12/24/accepted PY - 2012/2/21/entrez PY - 2012/2/22/pubmed PY - 2013/4/6/medline SP - e89 EP - 95 JF - Journal of the American Academy of Dermatology JO - J Am Acad Dermatol VL - 68 IS - 3 N2 - BACKGROUND: 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). OBJECTIVE: Development of reliable assays for the detection of anti-Col VII-NC1 antibodies. METHODS: 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). RESULTS: 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. LIMITATIONS: The novel assays were not evaluated prospectively and their use in monitoring serum levels during the disease course was not tested. CONCLUSION: 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. SN - 1097-6787 UR - https://www.unboundmedicine.com/medline/citation/22341608/Sensitive_and_specific_assays_for_routine_serological_diagnosis_of_epidermolysis_bullosa_acquisita_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0190-9622(12)00010-2 DB - PRIME DP - Unbound Medicine ER -