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Lupus anticoagulant

Order number Description Quantity Delivery time CE
CO 023-002 Anti-Cardiolipin IgG/IgM ELISA 3 plates 10-15 days

Anti-Cardiolipin IgG/IgM ELISA

For the detection and semi-quantitation of anti-cardiolipin antibodies in individuals with systemic lupus erythematosus (SLE) and lupus-like disorders (anti-phospholipid syndrome).

The test is performed as an indirect ELISA. Diluted serum samples, calibrator sera, and controls are incubated in cardiolipin coated microwells, allowing aCL antibodies present in the samples to react with the immobilized antigen. After the removal of unbound serum proteins by washing, antibodies specific for human IgG or IgM labeled with horseradish peroxidase (HRP) are added forming complexes with the cardiolipin bound antibodies. Two enzymeconjugated
antibody solutions are provided, one specific for human IgG antibodies and one specific for human IgM antibodies. The concentration of IgG aCL antibodies and IgM aCL antibodies must be determined separately. Following another washing step, the bound enzyme-antibody conjugate is assayed by the addition of  tetramethylbenzidine (TMB) and hydrogen peroxide (H2O2) as the chromogenic substrate. Color develops in the wells at an intensity proportional to the serum concentration of aCL antibodies.

Results are obtained by reading the O.D. (optical density or absorbance) of each well with a spectrophotometer. Calibrator sera are provided, with the IgG and IgM aCL concentrations expressed in GPL or MPL units, respectively. These units are traceable to the internationally recognized preparations of the Phospholipid Standardization Laboratory, University of Louisville. The user has the option of running either a single point calibrator or a four-point calibration curve. For single point calibration, dividing the concentration value of the
calibrator sera by the O.D. value of the calibrator provides a conversion factor (one for IgG and one for IgM aCL). The O.D. values of all the other samples are multiplied by the conversion factors to obtain IgG and IgM aCL antibody concentrations in standard units. One GPL unit is equivalent to 1 μg/mL of an affinity purified standard IgG sample, and one MPL unit is equivalent to 1 μg/mL of an affinity purified standard IgM sample.12 For multipoint calibration, perform a linear regression analysis with calibrator values against calibrator O.D.s. Control and patient results are determined from the calibration curve.



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