Abstract
The presence of antineutrophil cytoplasmic autoantibodies with specificity for proteinase 3 (PR3-ANCA) usually is detected by enzyme-linked immunosorbent assay (ELISA) with purified PR3 as a substrate. We studied the technical performance of direct and capture ELISA using a recombinant proteolytically inactive form of PR3 produced in the baculovirus expression system for the detection of PR3-ANCA in 114 patients with systemic vasculitis at diagnosis. We found that ELISA using recombinant PR3 produced in insect cells is a promising alternative for ELISA with native PR3. We found a correlation between tests using recombinant or native PR3, as well as correlation of the ELISA results with ANCA titers measured by the indirect immunofluorescence technique. However, the specificity for ANCA-associated vasculitis of ELISA with recombinant PR3 was lower than ELISA using native PR3. Compared with the direct assay, capture ELISA is a more sensitive method for PR3-ANCA detection, with both native and recombinant PR3, and its results depend on the monoclonal antibody used to capture the antigen.
Original language | English |
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Pages (from-to) | 586-595 |
Number of pages | 10 |
Journal | American Journal of Clinical Pathology |
Volume | 120 |
Issue number | 4 |
DOIs | |
Publication status | Published - Oct-2003 |
Keywords
- recombinant PR3
- proteinase 3
- PR3-ANCA
- antineutrophil cytoplasmic antibodies
- systemic vasculitis
- ELISA
- enzyme-linked immunosorbent assay
- ANTINEUTROPHIL CYTOPLASMIC ANTIBODIES
- WEGENERS-GRANULOMATOSIS
- CRESCENTIC GLOMERULONEPHRITIS
- MONOCLONAL-ANTIBODIES
- DISEASE-ACTIVITY
- CAPTURE-ELISA
- C-ANCA
- PR3
- AUTOANTIGEN
- EXPRESSION