Human Cardiolipin IgM ELISA Kit

Sample

Human serum or plasma (EDTA, citrate, heparin).

Species Reactivity

Human

Detection Method

ELISA

Intended Use

Enzyme immunoassay for the qualitative and quantitative determination of IgM antibodies against cardiolipin.

Contents of Kit

1. Microtiter Plate: coated with CL/ß2-GP1 and hermetically packed in a foil laminate pouch together with a desiccant bag. The plate consists of 12 strips, each of which can be broken into 8 individual wells.
2. Enzyme Conjugate IgM: 14 mL, ready-to-use, green colored. Buffered solution containing stabilising protein, methylisothiazolone and bromonitrodioxane.
3. Calibrator A-F: 2.0 mL each, 0 - 3.0 - 8.0 - 18 - 45 and 100 MPL-U CL/ß2-GP1 IgM / mL, ready-to-use, gradually blue colored. Contain TBS, BSA, Tween and Na-azide.
4. Negative and Positive Control: 2.0 mL each, ready-to-use, green and red colored, respectively. Contain TBS, BSA, Tween and Na-azide.
5. Sample Diluent: 100 mL, ready-to-use, orange colored. Contains Tris-buffered saline (TBS), bovine serum albumin (BSA), Tween and Na-azide.
6. TMB Substrate Solution: 14 mL, ready-to-use, colorless. Contains a buffered solution of TMB and H₂O₂. Contained in a vial impermeable to light.
7. Wash Buffer: 100 mL, 10× concentrate, blue colored. Contains TBS, Tween and bromonitrodioxane.
8. TMB Stop Solution (0.2 M H₂SO₄): 14 mL, colorless, ready-to-use. Caution: sulfuric acid is corrosive.
9. Instructions for Use.
10. Lot-specific certificate of analysis.

Storage

Store kit at 2 - 8°C, do not freeze. It is stable up to the expiry date stated on the label of the box. Do not use kit beyond its expiry date.

Performance Characteristics

1. Standardisation
The test is standardised with a purified serum preparation containing IgM antibodies specifically directed at CL/ß2-GP1. This preparation is calibrated against a set of commercially available, gradually positive sera ("Harris sera"; Louisville APL Diagnostics Inc., Louisville, KY, USA). The degree of sample reactivity is measured in MPL units (MPL-U CL/ß2-GP1 IgM / mL). 1 MPL-U/mL corresponds to the antigen binding capacity of a 1 μg/mL solution of IgM antibody affinity-purified from standard serum.
2. Analytical specificity
The test allows the specific determination of human IgM antibodies directed against CL/ß2-GP1. Interference with anticoagulants (EDTA, Citrat, Heparin) in samples has been tested and no interference effects have been observed.
3. Detection limit (analytical sensitivity)
The detection limit is defined as that concentration of analyte that corresponds to the mean absorbance of Sample Diluent plus 3-fold standard deviation (s). It was determined as < 0,5 MPL-U CL/ß2-GP1 IgM per mL sample (n = 24).
Recommended measuring range: 2 - 100 MPL-U CL/ß2-GP1 IgM per mL sample.
Specificity: 98%
Sensitivity: 100%

Precision

For the assessment of the test precision, the variability of results under the following conditions was determined:
a. within 1 assay and between 3 assays,
b. between 3 operators and
c. between 2 kit lots
a. Intra- and inter-assay variability (n = 24 and 72, respectively)

b. Operator to operator variability (n = 12)

c. Variability between 2 kit lots (n = 6)

General Description

Cardiolipin IgM ELISA is an enzyme-linked immunosorbent assay (ELISA) intended for the quantitative or qualitative determination of IgM class antibodies directed against Cardiolipin in human serum or plasma samples. Its function is the aid to diagnosis of anti-phospholipid syndrome (APS) and APS associated with systemic lupus erythematosus (SLE).