A promt diagnosis is essential for patients vasculitis or other inflammatory disorders. For ANCA positive samples which are negative for PR3-ANCA and MPO-ANCA an extended analysis is necessary. Wieslab? ANCA panel kit allows screening of six other factors that have been proposed to be involved in relevant disorders.
Intended use
The Wieslab? ANCA panel test kit is an enzyme-linked immunosorbent assay (ELISA) for qualitative detection of IgG antibodies to azurocidin, BPI, cathepsin G, elastase, lactoferrin, and lysozyme in human sera. The assay is used to detect antibodies in a single serum specimen. The test is useful as an aid in screening for specificity of ANCA IIF positive samples which are negative for PR3-ANCA and MPO-ANCA, or for detection of ANCA in inflammatory bowel diseases, liver diseases etc. One patient`s sample can be tested for all six antigens on twelve occasions. The analysis should be performed by trained laboratory professionals.
FOR IN VITRO DIAGNOSTIC USE.
Background
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 investigation of systemic vasculitis.
The first method to detect ANCA was indirect immunofluorescence (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.
In systemic vasculitis the two most important antigens are proteinase 3 and myeloperoxidase. In inflammatory conditions like rheumatoid arthritis, autoimmune hepatitis or inflammatory bowel disease however, many other antigens have been proposed to be involved. The most important being azurocidin, BPI, (BPI=Bactericidal permeability increasing protein), cathepsin G, elastase, lactoferrin and lysozyme.
Technical information
The wells of a microtitre strips are coated with purified ANCA antigens. 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 antibodies to human IgG binds to the antibodies in the wells in this 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 in terms of absorbance (optical density (OD)).
