Intended use

Enzyme immunoassay for the detection of autoantibodies IgM against Cardiolipin (CL).

Background

The anti-phospholipid syndrome (APS) is an autoimmune disorder which can comprise clinical conditions such as venous and arterial thrombosis, thrombocytopenia, myocardial infarction, recurrent spontaneous abortion and neurological complications.

Cardiolipin (CL) is the most common, negatively charged, acid phospholipid. Autoantibodies correlated with APS are directed not only against CL and similar phospholipides but also against phospholipid/protein complexes. ?2-glycoprotein 1 (?2-GP-1; = apolipoprotein H) has been identified as such a natural and essential co-antigen for CL-autoantibodies. The occurrence of CL/?2-GP-1 autoantibodies is associated with a tendency towards thrombosis. Besides this diagnostic significance, these antibodies are believed to be directly involved in the pathogenesis of APS.

Technical information

The EULISA Cardiolipin IgM is intended for the quantitative or qualitative determination of IgM antibodies in human serum, directed against CL/?2-GP-1. The immobilised antigen is a combination of CL, isolated from bovine heart, and a highly purified preparation of native ?2-GP-1. The test is fast (incubation time 30 - 30 - 30 minutes) and flexible (break apart wells, ready-to-use reagents). Six calibrators allow quantitative measurements; a negative and a positive control are used to check the assay performance.

Kit components and storage of reagents

Contents of the kit

1. 1 microwell plate, coated with CL/?2-GP-1 and hermetically packed in a foil laminate pouch together with a desiccant bag.

2. Sample buffer, 100 mL, ready-to-use, orange coloured.

3. Wash buffer, 100 mL, 10 x-concentrate, blue coloured.

4. 6 calibrators, 2.0 mL each: 0, 3.0, 8.0, 18, 45 and 100 MPL-U CL/?2-GP-1 antibodies (IgM)/mL, ready-to-use, gradually blue coloured.

5. Negative and positive control, 2.0 mL each, ready-to-use, green and red coloured, respectively.

6. Anti-human IgM HRP conjugate, 14 mL, ready-to-use, red coloured.

7. Substrate solution, 14 mL, ready-to-use, colourless.

8. Stop solution (0.5 M H2SO4), 14 mL, colourless, ready-to-use.

9. Directions for use

10. Lot-specific certificate of analysis

All kit components are stored at 2 - 8°C.