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ED-PR3 IU


 

Intended use
The  Wieslab® PR3-ANCA test kit is an enzyme-linked immunosorbent assay (ELISA) for detection and quantitation of IgG antibodies to proteinase 3 (PR3) 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 Wegener’s granulomatosis. The analysis should be performed by trained laboratory professionals.
FOR IN VITRO DIAGNOSTIC USE.

Principle of the Wieslab® PR3-ANCA
The wells of the microtitre plate are coated with purified proteinase 3. 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)). The absorbance is then calculated against a calibrator curve and the results are given in IU/mL adapted to the AF-CDC international standard for PR3.

Kit components and storage of reagents
- One frame with 96 wells (red coloured) coated with proteinase 3 with lid sealed in a foil pack with a dry pack.
- 1.5 mL negative control (NC) containing human serum in diluent.
- 1.5 mL positive  control (PC) containing human serum in diluent.
- 13 mL conjugate containing alkaline phosphatase-labelled antibodies to human IgG (blue colour).
- 2 x 32 mL Diluent (Dil) containing PBS (red colour).
- 13 mL Substrate pNPP.
- 13 mL Stop solution
- 30 mL wash solution 30x concentrated.
- Six calibrators (five calibrators, Cal 2-6, containing human serum) in diluent. 1.5 mL Cal 1 =
0 IU/mL, 1.5 mL Cal 2 = 2 IU/mL, 1.5 mL Cal 3 = 10 IU/mL, 1.5 mL Cal 4 = 30 IU/mL, 1.5 mL Cal 5 = 100 IU/mL, 1.5 mL Cal 6 = 200 IU/mL.

References
1. van der Woude FJ, et al. Autoantibodies against neutrophils and monocytes: Tool for diagnosis and marker of disease activity in Wegener´s granulomatosis. Lancet 1985, i 425-429.

2. Wiik A, Rasmussen N, Wieslander J. Methods to detect autoantibodies to neutrophilic granulocytes, in Van Venrooij WJ, Maini RN (eds): Manual of biological markers of disease. Kluwer Academic Publishers 1993, A9, 1-14.

3. Rasmussen N, Sjölin C, Isaksson B, Bygren P, Wieslander J. An ELISA for the detection of anti-neutrophil cytoplasm antibodies (ANCA). J Immunol Methods 1990, 127, 139-145.

4. Segelmark M, et al. How and why should we detect ANCA ? Clin Exp Rheumatol 2000,  18, 629-635.

5. Jenne DE, Tschopp J, Lüdemann J, Utecht B, Gross WL. Wegeners autoantigen decoded. Nature 1990, 346, 520.

6. Wieslander J, Wiik A. ANCA antigens: Proteinase 3, in van Venrooij W, Maini R (eds): Manual of Biological Markers of Disease. Kluwer Academic Publishers 1994, 1-9.

7. Falk RJ, Jennette JC. Anti-neutrophil cytoplasmic autoantibodies with specificity for myeloperoxidase in patients with systemic vasculitis and idiopathic necrotizing and crescentic glomerulonephritis. N Engl J Med 1988, 318, 1651-1657.

8. Wieslander J, Wiik A. ANCA antigens: Myeloperoxidase, in van Venrooij W, Maini R (eds): Manual of Biological Markers of Disease. Kluwer Academic Publishers 1994, 1-9.

9. Kamech L, Harper L, Savage C. ANCA - Positive Vasculitis. I. Am Soc Nephrol 2002 43 1953 - 1960.

10. Segelmark M, Elzouki AN, Wieslander J, Eriksson S. Heterozygosity for the alpha 1-antitrypsin PiZ gene affects the outcome of PR3-ANCA positive vasculitis. Kidney Int 1995, 48, 844-850.

11. Boomsma MM, Stegeman CA, van der Leij MJ, et al. Prediction of relapses in Wegener`s granulomatosis by measurement of ANCA levels: a prospective study. Arthritis Rheum, 2000, 43
2025 - 2033.

12. Savige J, et al.. ANCA and associated diseases: A review of the clinical and laboratory features. Kidney Int 2000, 57, 846-862.

13. Savige J, Gills D, Benson E et al. International Consensus Statement on Testing and Reporting of ANCA. Am J Clin Pathol 1999, 111, 507-513.

14. Won H, et al. Serial ANCA titers. Useful tool for the prevention of relapses in ANCA associated vasculitis. Kidney Int, 2003, 63, 1079-1085.

15. Neumann I, et al. Glomerular immune deposits are associated with increased proteinuria in patients with ANCA-associated crescentic nephritis. Nephrol Dial Transplant 2003, 18, 524-531.

16. Selga D. Diagnostic and prognostic aspects of anti-neutrophil cytoplasmatic antibodies in systemic vasculitis. Doctoral Dissertation, Department of Nephrology Clinical Science, Lund, 2007.

17.Csernok E, Ahlquist D, Ullrich S, Gross W.L. A critical evaluation of commercial immunoassays for antineutrophil

 ED-PR3 IU

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