Anti-Cardiolipin (aCL) Assays

Anti-Cardiolipin Autoimmune EIA Overview

Anti-Cardiolipin (aCL) Testing

The aCL assay serves to assist in the diagnosis of antiphospholipid syndrome. This diagnosis has important implications for the treatment and prediction of recurrent thrombosis and recurrent miscarriage.

The Bio-Rad Anti-Cardiolipin Test Kits are enzyme-linked immunosorbent assays (ELISA) for the detection and semi-quantitative determination of aCL IgG, IgM, or IgA antibodies in human serum or plasma. The results are reported in GPL, MPL, and APL units that are directly traceable to an internationally recognized reference preparation. The Bio-Rad Anti-Cardiolipin Test Kits are intended to be used by clinical (hospital and reference) laboratories.

Product Highlights

• Specific determinations of IgG, IgM, and IgA aCL antibodies
• Flexible multi-point or single-point calibration options
• Better regression analysis with distinct calibrators
• Built-in quality control with undiluted controls and calibrators

Anti-Cardiolipin Autoimmune EIA

Pre-Test Setup Instructions

Step 1. Bring all reagents to room temperature (18–26°C). Step 2. Prepare the Wash Solution.
Step 3. Make 1:50 working solutions by diluting 10 µL of patient's serum positive/negative controls, and 3 calibrators in 490 µL of diluent.

Test Procedure

Step 1. Apply 100 µL diluted patient samples, controls, and calibrator to wells. Step 2. Incubate wells for 30–40 minutes at room temperature (18–26°C).
Step 3. Wash wells 4x with ~250 µL of wash solution.Remove all liquid before proceeding.
Step 4. Apply 100 µL HRP conjugate to all wells. Discard excess transferred HRP after use.
Step 5. Incubate wells for 30–40 minutes at room temperature (18–26°C). Step 6. Wash wells 4x with ~250 µL of wash solution. Remove all liquid before proceeding. Step 7. Develop color by adding 100 µL substrate to each well. Discard excess transferred substrate after use.
Step 8. Incubate for 30–40 minutes at room temperature (18–26°C) Step 9. Stop color development by adding 100 µL of stop solution to each well. Step 10. Read results within 30 minutes with an EIA reader set to 450 nm.

Anti-Cardiolipin Autoimmune EIA

aPL Testing Algorithm: Antiphospholipid Antibody Screening Procedure

* Screen for each isotype. Polyvalent screen not recommended.

APS = Antiphospholipid syndrome
aCL = Anti-cardiolipin
aPS = Anti-phosphatidylserine
β2 GPI = β-2 glycoprotein 1 (cofactor)
aPT = Anti-prothrombin

Cut-Off Value

• aCL IgG: <23 GPL units
• aCL IgA: <22 APL units
• aCL IgM: <11 MPL units