A promt diagnosis is essential for patients with medical conditions such as granulomatosis with polyangiitis (Wegener's granulomatosis) or microscopic polyangiitis. Fast and simultaneous detection of Proteinase 3 (PR3-ANCA) and Myeloperoxidase (MPO-ANCA) is possible with Wieslab® ANCA screen kit.
The Wieslab® ANCA screening test kit is an enzyme-linked immunosorbent assay (ELISA) for qualitative detection of IgG antibodies to proteinase 3 and myeloperoxidase (MPO) in human sera. The assay is used to detect antibodies in a single serum specimen. The results of the assay are to be used as an aid to the diagnosis of systemic vasculitis, especially granulomatosis with polyangiitis (Wegener’s granulomatosis (WG)) and microscopic polyangiitis (MP). The analysis should be performed by trained laboratory professionals. FOR IN VITRO DIAGNOSTIC USE.
A positive result should always be confirmed by a semi-quantitative assay.
ANCAs (anti-neutrophil cytoplasmic antibodies) are a family of autoantibodies related to vasculitis and inflammatory disorders. Since 1985, when c-ANCA was shown to be related to Wegener’s granulomatosis, interest in ANCAs has steadily increased, and today these antibodies are considered to be major diagnostic tools for the diagnosis and follow up of systemic vasculitis.
The first method to detect ANCA was indirect immuno-fluorescence (IIF) performed on ethanol fixed granulocytes. This method yields two patterns, a cytoplasmic staining of the granulocyte denoting the presence of c-ANCAs, and a perinuclear staining denoting the presence of p-ANCAs. IIF was followed by ELISAs using the purified proteins.
The granulocyte is full of granules each with many different proteins. It was early shown that antibodies from systemic vasculitis patients bind to the alpha fraction containing the
azurophil granules. The most important proteins were proteinase 3 (PR3) and myeloperoxidase (MPO). Thus antibodies to proteinase 3 are termed PR3-ANCA, and antibodies to myeloperoxidase are termed MPO-ANCA.
Approximately 80-90% of WG patients manifest PR3-ANCA and 5-15% MPO-ANCA. One category of vasculitis is microscopic polyangiitis (MP). Most patients with active MP are characterised by positive ANCA test results, MPO-ANCA being more frequent than PR3-ANCA.
The wells of the microtiterstrip are coated with purified proteinase 3 and myeloperoxidase. During the first incubation, specific antibodies in diluted serum will bind to the antigen coating. The wells are then washed to remove unbound antibodies and other components. A conjugate of alkaline phosphatase labelled (goat) antibodies to human IgG binds to the antibodies in the wells in the second incubation. After a further washing step, detection of specific antibodies is obtained by incubation with substrate solution. The amount of bound antibodies correlates to the colour intensity and is measured as absorbance (optical density (OD)).