Human Cardiolipin IgM ELISA Kit

Sample

Human sera

Species Reactivity

Human

Detection Method

sELISA

Intended Use

The Cardiolipin Antibody Test System is an enzyme-linked immunosorbent assay (ELISA) designed for the semi-quantitative measurement of circulating IgM autoantibodies to cardiolipin. For research use only.

Contents of Kit

Each kit contains the following components in sufficient quantities to perform the number of tests indicated on packaging label.
1. Plate. 96 wells configured in twelve 1x8-well strips coated with Cardiolipin antigen from bovine heart. The strips are packaged in a strip holder and sealed in an envelope with desiccant.
2. Conjugate. Conjugated (horseradish peroxidase) goat anti-human IgM (chain specific). Ready to use. One, 15 ml vial with a white cap. Preservative added.
3. Positive Control (Human Serum). One, 0.35 mL vial with red cap. Preservative added.
4. Calibrator (Human Serum). One, 0.5 mL vial with a blue cap. Preservative added.
5. Negative Control (Human Serum). One, 0.35 mL vial with a green cap. Preservative added.
6. Sample diluent. One 30 mL bottle (green cap) containing Tween 20, bovine serum albumin and phosphate-buffered-saline, (ph 7.2 ± 0.2). Green solution, ready to use. Note: Shake Well Before Use. Preservative added.
7. TMB: One 15 mL amber bottle (amber cap) containing 3,3',5,5,'-tetramethylbenzidine (TMB). Ready to use. Contains DMSO ≤15% (w).
8. Stop Solution: One 15 mL bottle (red cap) containing 1M H₂SO₄, 0.7M HCI. Ready to use.
9. Wash Buffer concentrate (10X): dilute 1 part concentrate + 9 parts deionized or distilled water. One 100 mL bottle (clear cap) containing a 10X concentrated phosphate-buffered-saline and Tween-20 solution (clear solution). Contains preservative. Note: 1X solution will have a pH of 7.2 ± 0.2.
The following components are not kit lot number dependent and may be used interchangeably with this ELISA assay: TMB, Stop Solution.
Note: Kit also contains:
1. Component list containing lot specific information is inside the kit box.
2. Package insert providing instructions for use.

Storage

1. Store the unopened kit at 2°C and 8°C.
2. Coated microwell strips: Store between 2°C and 8°C. Extra strips should be immediately resealed with desiccant and returned to proper storage. Strips are stable for 60 days after the envelope has been opened and properly resealed and the indicator strip on the desiccant pouch remains blue.
3. Conjugate: Store between 2°C and 8°C. DO NOT FREEZE.
4. Calibrator, Positive Control and Negative Control: Store between 2°C and 8°C.
5. TMB: Store between 2°C and 8°C.
6. Wash Buffer concentrate (10X): Store between 2°C and 25°C. Diluted wash buffer (1X) is stable at room temperature (20°C - 25°C) for up to 7 days or for 30 days between 2°C and 8°C.
7. Sample Diluent: Store between 2°C and 8°C.
8. Stop Solution: Store at 2°C and 25°C.

Performance Characteristics

Comparative Study An in-house comparative study was performed to demonstrate the equivalence of the Cardiolipin IgM ELISA (DEIA1709) test system to another commercially available Cardiolipin IgM ELISA test system. Performance was evaluated using 260
specimens as described in Table 1 below. The results of the investigation have been summarized in Table 2 below.
Table 1. Summary of Clinical Specimens

Table 2. Calculation of Relative Sensitivity, Specificity, and Agreement

Relative Sensitivity = 44/47 = 93.6%
95% Confidence Interval* = 86.6 to 100%
Relative Specificity = 163/213 = 76.5%
95% Confidence Interval* = 70.8 to 82.2%
Relative Agreement = 207/ 260 = 79.6%
95% Confidence Interval* = 74.7 to 84.5%
* 95% confidence intervals calculated using the exact method.

General Description

Autoantibodies directed against phopholipids, and anti-cardiolipin (aCL) in particular, have been associated with recurrent thrombosis, thrombocytopenia, and spontaneous abortions (1, 2, 3). aCL is observed in patients with systemic lupus erythematosus, in patients with other connective tissue disease (4), in individuals undergoing chlorpromazine treatment (5), as well as in persons who do not have chronic illness.