Diagnostics of autoimmune blistering disorders: an experience of a single tertiary referral centre.

Autoimmune blistering disorders (AIBD) include a heterogeneous group of diseases characterized by the presence of autoantibodies against the structural antigens of the skin and mucous membranes. The gold standard of AIBD diagnostics is the detection of in vivo bound IgG/IgA and/or complement component 3 in the direct immunofluorescence of a perilesional biopsy. Various immunological techniques such as indirect immunofluorescence of different tissue substrates including monkey oesophagus, salt split skin, recombinant proteins of epidermis and basement membrane zone as well as ELISA systems and immunoblotting are used to characterize target antigens. Proper and early diagnosis is crucial for both treatment and prognosis since some AIBD may be associated with a malignant neoplasm.

Case Report: Pemphigoid Nodularis-Five Patients With Many Years of Follow-Up and Review of the Literature.

Pemphigoid nodularis is a rare form of pemphigoid that joins the clinical picture of prurigo nodularis and the immunological features of bullous pemphigoid, which is therapeutically challenging. Here, we analyze five female patients with a long-lasting course of nodular pemphigoid in terms of clinical and immunological characteristics and therapy. All the patients fulfilled clinical and immunological criteria of nodular pemphigoid. We applied numerous techniques allowing the proper diagnosis: direct and indirect immunofluorescence, salt split skin, ELISA, BIOCHIP, and fluorescence overlay antigen mapping using laser scanning confocal microscopy. Our study showed that 4 of 5 patients fulfilled the clinical and immunological criteria of nodular bullous pemphigoid. Two out of 4 patients presented exclusively nodular lesions; in the other two patients, blisters and erythematous lesions preceded prurigo-like lesions by a few years. The remaining patient had clinical and immunological criteria of nodular mucous membrane pemphigoid, presenting oral erosions, scarring conjunctivitis, and numerous disseminated nodules on the skin. All the patients were treated with multiple medicines; however, it was observed that the use of clobetasol propionate on the entire body plus antidepressants best controlled the disease. Pemphigoid nodularis mainly occurs in elderly women. In cases with coexisting psychological problems, antidepressants should be considered as an important complementary therapy to the basic one with clobetasol propionate.

The use of BIOCHIP mosaics in diagnostics of bullous pemphigoid: Evaluation and comparison to conventional multistep procedures.

BACKGROUND: Bullous pemphigoid (BP) is an autoimmune blistering disease associated with autoantibodies against BP180 and/or BP230 antigens. The immunoassays available for serological diagnostics include indirect immunofluorescence (IIF) on monkey esophagus (ME), salt-split skin (SSS), and enzyme-linked immunosorbent assay (ELISA) for BP180-NC16a and BP230. Only a few studies validated innovative BIOCHIP mosaic, but none compared agreement between BIOCHIP substrates with conventional methods separately. METHODS: We evaluated the agreement between BIOCHIP and conventional methods and assessed sensitivity and specificity in BP diagnosis. The study comprised 51 BP patients and 39 controls. RESULTS: Analysis showed very good agreement between BIOCHIP-SSS vs classic IIF-SSS (0.933, P < 0.001) and for BIOCHIP-BP180-NC16a vs ELISA-BP180-NC16a (0.933, P < 0.001). A good strength of agreement between BIOCHIP-ME vs classic IIF-ME was observed (0.694, P < 0.001) similar to BIOCHIP-BP230 vs ELISA-BP230 (0.793, P < 0.001). BIOCHIP-ME sensitivity was 51.0%, whereas IIF-ME was 76.5%. Epidermal reaction on BIOCHIP-SSS was found in 94.1% of BP patients and in all patients on IIF-SSS (sensitivity 100%). BIOCHIP-BP180-NC16a sensitivity was lower than in ELISA-BP180-NC16a (76.5% vs 82.4%). BP230 sensitivity of both methods was similar (45.1% vs 43.1%). The specificity for all antigens was 100%. CONCLUSION: BIOCHIP mosaic is a useful method presenting satisfactory agreement with conventional immunoassays.

The use of BIOCHIP technique in diagnosis of different types of pemphigus: Vulgaris and foliaceus.

BACKGROUND: Pemphigus is a rare, autoimmune blistering disease characterized by autoantibodies against desmoglein 3 (Dsg3) and 1 (Dsg1) with mucosal and/or skin involvement. Main types of pemphigus include mucosal pemphigus vulgaris (m-PV), mucocutaneus pemphigus vulgaris (mc-PV) and pemphigus foliaceus (PF) determined by clinical picture, positive direct and indirect immunofluorescence, and enzyme-linked immunosorbent assay (ELISA). METHODS: We evaluated the sensitivity and specificity of a novel multi-substrate immunofluorescence technique called BIOCHIP in the diagnosis of main types of pemphigus. Additionally, we tested agreement between BIOCHIP-Dsg1 and ELISA-Dsg1 in differentiation pemphigus vulgaris subtypes. The study comprised 35 patients with pemphigus: 14 patients with PF, 21 with PV (13 with m-PV and 8 with mc-PV) and 48 controls. RESULTS: The intercellular staining on monkey esophagus substrate in BIOCHIP was observed in 23/35 pemphigus in total (sensitivity 65.7%), 17/21 PV (sensitivity 81.0%), 10/13 m-PV (sensitivity 76,9%), 7/8 mc-PV (sensitivity 87.5%) and 6/14 PF (sensitivity 42.9%), but not in 48 controls. Dsg3 positive staining in BIOCHIP was observed in 21/21 PV (sensitivity 100%), 13/13 m-PV (sensitivity 100%), 8/8 mc-PV (sensitivity 100%), whereas Dsg3 was negative in all 14 PF sera. Dsg1 reactivity was detected in 9/21 PV (sensitivity 42.8%), 2/13 m-PV (sensitivity 15,4%), 7/8 mc-PV (sensitivity 87.5%) and 13/14 PF (sensitivity 92.9%). All 48 controls were negative for both Dsg3 and Dsg1. An excellent agreement for BIOCHIP-Dsg1 and ELISA-Dsg1 for m-PV and mc-PV was found, which reflect k values of 1.0 and 0.91, respectively. CONCLUSION: BIOCHIP technique is a useful method for pemphigus diagnostics and differentiation between its subtypes: m-PV, mc-PV and PF.