The data and the results obtained at the above symposium show that the flow cytometric method closely correlates with the standardized aCL bench ELISA and with the APhL ELISA kit. The aCL/aPS FACS kit is comparable in sensitivity to the APhL ELISA kit and standard aCL bench ELISA, but the aCL/aPS FACS kit is more specific than the standard anticardiolipin assay (particularly for the determination of aPS). In summary, the aCL/aPS FACS kit enables rapid (total run time < or = 1 hr) and simultaneous determination of aCL (aCL) and anti-PS antibodies of the IgG and IgM classes and combines good sensitivity and specificity in a single assay. The method is reproducible (intraassay variations < 5%). Furthermore, it is easy to transform into a partially or fully mechanized process and is well suited for laboratories that test large numbers of samples daily. The assay promises to be useful not only for detecting positive APS sera, but also in evaluating the significance of phospholipid specificity and antibody isotypes in patients with APS.

American Clinical Laboratory

A flow cytometric assay for the detection of antiphospholipid antibodies

Author: Silva, Lalith K. ; Harris, E. Nigel ; Pierangeli, Silvia S.

A procedure for detection of antiphospholipid antibodies using flow cytometry is described.Using beads of different sizes antibodies to cardiolipin (CL) and phosphatidylserine (PS) were detected simultaneously in a single tube.The presence of IgG or IgM and their reactivities to CL and PS were analyzed.The procedure for performance of the assay, its sensitivity and its specificity are discussed.

Medicina & Laboratorio

Síndrome antifosfolípido: mecanismos patogénicos, diagnóstico y tratamiento

Author: Silvia S Pierangeli ; Héctor R Pierangeli

Antiphospholipid antibodies are associated with thrombosis and pregnancy losses in patients with the antiphospholipid syndrome. Several pathogenic mechanisms have been described to explain clinical manifestations produced by antiphospholipid antibodies. This review addresses current modalities of treatment as well as an update on tests used for the confirmation of the diagnosis of antiphospholipid syndrome. The anticardiolipin test has been widely utilized by physicians since the mid-1980’s for diagnosing patients with antiphospholipid syndrome. Establishment of this diagnosis has enabled effective management of patients with recurrent thrombosis and recurrent pregnancy losses. The test was first established in 1983 as a radioimmunoassay and soon thereafter converted into an enzyme-linked immunosorbent assay (ELISA). The other test commonly used in the diagnosis of antiphosphoplipid syndrome is the lupus anticoagulant test. The anticardiolipin test by ELISA is sensitive for the diagnosis of antiphospholipid syndrome but lacks speci- ficity. On the other hand, the lupus anticoagulant assay, although more specific is not as sensitive as the anticardiolipin ELISA assay. More specific tests are now available such as the anti-s2 glycoprotein I (anti-s2GPI), the anti-prothrombin (anti-PT) assay and the APhL ELISA test that utilizes negatively charged phospholipids instead of cardiolipin to coat the plates. This article discusses in detail the clinical value of the above-mentioned tests, technical problems associated with them, the current laboratory classification criteria for diagnosis of antiphospholipid syndrome and possible new and better assays that will be available in the near future for diagnosis of anti- phospholipid syndrome.

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