ED-CAP-MPO IU

Intended use
The Wieslab® Capture MPO-ANCA test kit is an enzyme-linked immunosorbent assay (ELISA) for detection and quantitation of IgG antibodies to myeloperoxidase (MPO) in human sera. The results of the assay are to be used as an aid to the diagnosis of microscopic polyangiitis (MP). The assay is intended for use in patients with signs and symptoms consistent with MP. It is not intended for screening a healthy population. The analysis should be performed by trained laboratory professionals.
FOR IN VITRO DIAGNOSTIC USE.

Principle of the Wieslab® Capture MPO-ANCA ELISA
The wells of the microtitre plate are coated with purified anti-MPO monoclonal antibody 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 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 MPO.

Kit components and storage of reagents
- One frame with 96 wells (green coloured) coated with monoclonal
anti-myeloperoxidase/myeloperoxidase 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 in PBS (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) serum in diluent.
1.5 mL Cal 1 = 0 IU/mL, 1.5 mLCal 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.
All reagents in the kit are ready for use except wash solution and should be stored at 2-8° C.
Remove only the number of wells needed for testing, resealing the aluminium package carefully.

References
1. 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.
Dordrecht, Kluwer Academic Publishers; 1993:A9,1-14.

2. Rasmussen N, Sjolin 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.

3. Wieslander J. How are anti neutrophil cytoplasmic autoantibodies detected?
Am J  Kidney Dis 1991;18:154-158.

4. Jenne DE, Tschopp J, Ludemann J, Utecht B, Gross WL. Wegeners autoantigen  decoded.
Nature 1990;346:520.

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

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

7. Jennette C, Falk R. Small vessels vasculitis.
N Engl J Med 1997, 337, 1512-1525.

8. 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.

9. Cohen Tervaert JW, Huitema MG, Hene RJ, Sluiter WJ, The T, van der Hem GK et  al. Prevention of relapses