Abstract
BackgroundVarious antigen-specific immunoassays are available for the serological diagnosis of autoimmune bullous diseases. However, a spectrum of different tissue-based and monovalent antigen-specific assays is required to establish the diagnosis. BIOCHIP mosaics consisting of different antigen substrates allow polyvalent immunofluorescence (IF) tests and provide antibody profiles in a single incubation.MethodsSlides for indirect IF were prepared, containing BIOCHIPS with the following test substrates in each reaction field: monkey esophagus, primate salt-split skin, antigen dots of tetrameric BP180-NC16A as well as desmoglein 1-, desmoglein 3-, and BP230gC-expressing human HEK293 cells. This BIOCHIP mosaic was probed using a large panel of sera from patients with pemphigus vulgaris (PV, n = 65), pemphigus foliaceus (PF, n = 50), bullous pemphigoid (BP, n = 42), and non-inflammatory skin diseases (n = 97) as well as from healthy blood donors (n = 100). Furthermore, to evaluate the usability in routine diagnostics, 454 consecutive sera from patients with suspected immunobullous disorders were prospectively analyzed in parallel using a) the IF BIOCHIP mosaic and b) a panel of single antibody assays as commonly used by specialized centers.ResultsUsing the BIOCHIP mosaic, sensitivities of the desmoglein 1-, desmoglein 3-, and NC16A-specific substrates were 90%, 98.5% and 100%, respectively. BP230 was recognized by 54% of the BP sera. Specificities ranged from 98.2% to 100% for all substrates. In the prospective study, a high agreement was found between the results obtained by the BIOCHIP mosaic and the single test panel for the diagnosis of BP, PV, PF, and sera without serum autoantibodies (Cohen’s κ between 0.88 and 0.97).ConclusionsThe BIOCHIP mosaic contains sensitive and specific substrates for the indirect IF diagnosis of BP, PF, and PV. Its diagnostic accuracy is comparable with the conventional multi-step approach. The highly standardized and practical BIOCHIP mosaic will facilitate the serological diagnosis of autoimmune blistering diseases.
Highlights
Various antigen-specific immunoassays are available for the serological diagnosis of autoimmune bullous diseases
Intercellular epithelial staining on monkey esophagus by indirect IF was observed in 65 pemphigus vulgaris (PV) and 49 pemphigus foliaceus (PF) sera resulting in sensitivities of 100% and 98.0%, respectively
A BIOCHIP mosaic was composed allowing the simultaneous detection of autoantibodies against the most frequent target antigens to facilitate the diagnosis of bullous skin diseases
Summary
Various antigen-specific immunoassays are available for the serological diagnosis of autoimmune bullous diseases. Autoimmune bullous disorders are characterized by autoantibodies against desmosomal proteins (in pemphigus), adhesion molecules of the dermal-epidermal junction (in pemphigoid diseases), and epidermal/ tissue transglutaminase (in dermatitis herpetiformis), respectively [1,2,3]. The most frequent autoimmune bullous diseases are bullous pemphigoid and pemphigus, with incidences varying considerably between geographical regions [4,5,6,7,8]. In central Europe, pemphigus is less frequent with incidences ranging from 0.6 to 6.8 new patients/ million/year [5,8,11], higher incidences can be found in Southeastern Europe, the Mediterranean region, Iran and the Jewish population [7,12]. Incidences of the other entities are below 1.0/million/year
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