Serological diagnosis of autoimmune bullous skin diseases: Prospective comparison of the BIOCHIP mosaic-based indirect immunofluorescence technique with the conventional multi-step single test strategy

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.

The 2016 Bowman Lecture Conjunctival curses: scarring conjunctivitis 30 years on

“…However, the prevalence of these antibodies has varied in relation to the sensitivity and specificity of the assay used and the control group. The lowest prevalence has been less than 1% in a large cohort of blood donors of unknown ages, 54 other publications have given values of 7.5%, 55 13%, 16% and 26% for ELISA and or immunoblots 56 although higher values have been reported. 57 Results for IIF have generally shown o5% positivity 55,56 although this was 19% (n = 32) in another study.…”

“…The lowest prevalence has been less than 1% in a large cohort of blood donors of unknown ages, 54 other publications have given values of 7.5%, 55 13%, 16% and 26% for ELISA and or immunoblots 56 although higher values have been reported. 57 Results for IIF have generally shown o5% positivity 55,56 although this was 19% (n = 32) in another study. 58 Autoreactive T cells recognising the basement membrane protein BP 180, are also present in healthy individuals expressing the HLA-DQB*0301 gene (although not in those who did not).…”

Clinical features and diagnosis of epidermolysis bullosa acquisita

“…In autoimmune blistering diseases, a BIOCHIP ® mosaic containing six substrates comprising monkey esophagus, salt-split skin, recombinant BP180 NC16A, and HEK293 cells transfected for expression of desmoglein 1, desmoglein 3, and BP230 is widely used in the dermatological community [136,[153][154][155][156].…”

Bullous pemphigoid resembling erythema gyratum repens – case report

“…As a result, it is possible to simultaneously test one sample for antibodies against several substrates, which shortens the testing procedure. The substrates used in the diagnostics of pemphigoid with the method include: monkey oesophagus, primate salt-split skin, and antigen dots of tetrameric BP180-NC16A and BP230gC-expressing cells [12].…”