Human cardiolipin as the antigen in an ELISA to detect anticardiolipin antibodies
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1Haematology Department, Royal Prince Alfred Hospital, Camperdown, NSW, Australia.
Insights
Using human cardiolipin in enzyme-linked immunoassays (ELISA) for anticardiolipin antibodies improves diagnostic specificity. This method enhances detection of antibodies in lupus anticoagulant positive patients.
Area of Science:
- Immunology
- Biochemistry
Background:
- Anticardiolipin antibodies are associated with autoimmune diseases, particularly lupus anticoagulant.
- Current enzyme-linked immunoassays (ELISA) often use bovine cardiolipin as the antigen.
Purpose of the Study:
- To evaluate the specificity and sensitivity of using human cardiolipin as an antigen in ELISA for detecting anticardiolipin antibodies.
- To compare the diagnostic performance of human versus bovine cardiolipin in patients with lupus anticoagulant.
Main Methods:
- Human cardiolipin was purified from cadaver heart tissue.
- An enzyme-linked immunoassay (ELISA) was developed using human cardiolipin as the antigen.
- Assays were performed on patient samples, including those positive for lupus anticoagulant.
- Results were compared with assays using bovine cardiolipin and sheep cardiolipin.
Main Results:
- A high correlation (r = 0.90) was observed between human and bovine cardiolipin assays.
- 88% of lupus anticoagulant positive patients had antibodies to human cardiolipin, compared to 75% for bovine cardiolipin.
- Six samples (13.3%) showed reactivity exclusively with the human antigen and not sheep cardiolipin.
- These exclusively reactive antibodies remained consistent across different human cardiolipin preparations.
Conclusions:
- Human cardiolipin offers increased specificity as a phospholipid antigen in ELISA for anticardiolipin antibodies.
- The use of human cardiolipin may improve the diagnostic accuracy for detecting specific anticardiolipin antibodies, particularly in lupus anticoagulant positive individuals.
Abstract:
Human cardiolipin was purified from cadaver heart tissue and used as the antigen in an enzyme-linked immunoassay (ELISA) for anticardiolipin antibodies. The correlation between assays using human and bovine cardiolipin was r = 0.90. Of the lupus anticoagulant positive patients 88% had antibodies to the human antigen and 75% had antibodies to the bovine antigen. Six of 45 samples contained antibodies which only reacted with the human antigen. All six samples remained positive when retested with a different human cardiolipin preparation and were negative when tested against a preparation of sheep cardiolipin. These studies suggest that there was increased specificity when human cardiolipin was used as the phospholipid antigen.