Cardiolipin IgM ELISA Kit

Sample

Saliva

Species Reactivity

Human

Intended Use

The Cardiolipin Antibody Test System is an enzyme-linked immunosorbent assay (ELISA) designed for the semiquantitative measurement of circulating IgM autoantibodies to cardiolipin.

Contents of Kit

1. [PLATE] Plate.
2. [CONJ] Conjugate.
3. [CONTROL+] Positive Control (Human Serum).
4. [CAL] Calibrator (Human Serum).
5. [CONTROL-] Negative Control (Human Serum).
6. [DILSPE] Sample diluent.
7. [SOLN TMB] TMB:
8. [SOLN STOP] Stop Solution:
9. [WASHBUF 10x] Wash Buffer concentrate (10x): dilute 1 part concentrate + 9 parts deionized or distilled water.

Storage

2°C to 8°C

Performance Characteristics

##Comparative Study##
An in-house comparative study was performed to demonstrate the equivalence of the Cardiolipin IgM ELISA 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.

##Reproducibility##
To evaluate both intra-assay and inter-assay reproducibility, six specimens were tested; eight replicates each, on each of three days. These results were then used to calculate mean unit values, standard deviations, and percent CV. Two of the specimens were strong positives, two were clearly negative, and two were near the assay cut off. The results of the study have been summarized below.
Table 3. Cardiolipin IgM ELISA ; Results of Precision Testing
Day 1: Intra-Assay Reproducibility

Day 2: Intra-Assay Reproducibility

Day 3: Intra-Assay Reproducibility

Intra-Assay Reproducibility; All Days Combined:


##Cross-Reactivity##
To investigate the potential for positive reactions due to cross-reactive antibodies, fourteen specimens which were reactive for various autoantibodies were tested on the anti-Cardiolipin test system. None of the fourteen were positive for antiCardiolipin IgM antibody. The results of this study indicate that the potential for a high degree of cross reactivity with such autoantibodies is not likely.

General Description

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

Reconstitution And 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 to 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.