Development of an ELISA for rapid detection of anti-type VII collagen autoantibodies in epidermolysis bullosa acquisita.
Epidermolysis bullosa acquisita (EBA) is an acquired blistering skin disease characterized by the presence of IgG autoantibodies to type VII collagen. EBA autoantibodies recognize four major immunodominant epitopes localized within the amino-terminal, noncollagenous (NC1) domain. In this study, we developed a rapid, quantitative enzyme-linked immunosorbent assay (ELISA) to detect autoantibody activity against the complete NC1 domain of type VII collagen with the use of an eukaryotic-expressed, recombinant human NC1 antigen. With the ELISA, we tested serum from patients with EBA (n = 24), bullous systemic lupus erythematosus (BSLE) (n = 3), bullous pemphigoid (n = 16), pemphigus (n = 11), and normal controls (n = 12). All EBA and BSLE serum, including four sera that were negative by indirect immunofluorescence, demonstrated reactivity with immobilized NC1 in the ELISA. In contrast, none of the sera from healthy control subjects or patients with unrelated blistering skin diseases reacted with NC1. The EBA sera also reacted with recombinant NC1 by immunoblot analysis but with less sensitivity. Thus, the newly developed ELISA using recombinant NC1 is a sensitive, specific assay and a useful tool for rapidly screening EBA and BSLE serum.
Chen
M
M
Department of Dermatology, Northwestern University Medical School, Chicago, Illinois 60611, USA.
Chan
L S
LS
Cai
X
X
O'Toole
E A
EA
Sample
J C
JC
Woodley
D T
DT
D001323:Autoantibodies / Q000032:analysis*; D003094:Collagen / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D016107:Epidermolysis Bullosa Acquisita / Q000097:blood / Q000276:immunology*; D006801:Humans; D008178:Lupus Erythematosus, Cutaneous / Q000097:blood / Q000276:immunology; D008180:Lupus Erythematosus, Systemic / Q000097:blood / Q000276:immunology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012872:Skin Diseases, Vesiculobullous / Q000097:blood / Q000276:immunology
D001323:Autoantibodies; D003094:Collagen