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
PY - 2013/3/1
Y1 - 2013/3/1
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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=84873708269&partnerID=8YFLogxK
U2 - 10.1016/j.jaad.2011.12.032
DO - 10.1016/j.jaad.2011.12.032
M3 - Journal articles
C2 - 22341608
AN - SCOPUS:84873708269
SN - 0190-9622
VL - 68
JO - Journal of the American Academy of Dermatology
JF - Journal of the American Academy of Dermatology
IS - 3
ER -