AUTOIMMUNE SKIN DISEASE

Instructions for use

The MESACUP Desmoglein TEST “Dsg3” is a semi-quantitative enzyme-linked immunosorbent assay (ELISA) for the detection of anti-Desmoglein3 antibodies in human serum.

Background:

Desmoglein 3 (Dsg3) is a desmosomal cadherin expressed in stratified squamous epithelia and the target antigen in pemphigus vulgaris (PV). PV is a fatal autoimmune blistering disease of the skin and mucous membrane and clinically characterized by flaccid blisters and widespread painful erosions. Histologically, the lesions of PV demonstrate intraepidermal blister formation due to loss of cell adhesion of keratinocytes. Anti-Dsg3 autoantibodies (IgG) are detected in sera from patients with PV and play a pathogenic role in blister formation in PV. Anti-Dsg3 IgG is not found in patients with pemphigus foliaceus (PF), another type of pemphigus. Amagai et al have produced a recombinant human Dsg3 (rDsg3) using a baculovirus expression system. rDsg3 is designed as a secreted protein consisting of the entire extracellular domain of Dsg3. rDsg3 is capable of immunoadsorbing pathogenic autoantibodies in PV patients' sera, preventing blister formation in a neonatal mouse model of pemphigus. These findings indicate that rDsg3 expresses most, if not all, conformational epitopes of the native antigens critical in the pathogenesis of pemphigus. Currently, diagnosis of pemphigus largely relies on immunofluorescence testing (IF) utilizing sectioned normal human skin or monkey esophagus. However, there are several disadvantages in the current immunofluorescence. (a) IF requires an experienced examiner and a subjective determination of antibody titer. (b) Antibody against any cell surface protein makes false positive. (c) Similar staining pattern in IF makes it difficult to distinguish between PV and PF. To overcome the problems of IF, ELISA has been developed using rDsg3 as a sensitive and specific assay for the diagnosis of pemphigus.

The MESACUP Desmogrein TEST “Dsg3” is for measuring anti Desmoglein 3 IgG autoantibodies with high sensitivity.

Principle:

MESACUP Desmoglein TEST ”Dsg3” measures anti-Dsg3 antibodies present in the serum by ELISA. Calibrators and patient sera are added to microwell coated with Dsg3, allowing anti-Dsg3 antibodies to react with the immobilized antigen (Sample incubation). After wash to remove any unbound serum proteins, horseradish peroxidase conjugated anti-human IgG monoclonal antibody is added and incubated (Conjugate incubation). Following another washing step, the peroxidase substrate is added and incubated for an additional period of time (Substrate incubation). Acid solution is then added to each well to terminate the enzyme reaction and to stabilize the color development. The assay can be quantified by measuring the reaction photometrically.

Size: 48 wells

Method: ELISA

Antigen: recombinant

Availability: all regions

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