Laboratory detection of the antiphospholipid syndrome via calibrated automated thrombography

Katrien Devreese; Kathelijne Peerlinck; Jef Arnout; Marc F. Hoylaerts

doi:10.1160/TH08-06-0393

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2009; 101(01): 185-196
DOI: 10.1160/TH08-06-0393

New Technologies, Diagnostic Tools and Drugs

Schattauer GmbH

Laboratory detection of the antiphospholipid syndrome via calibrated automated thrombography

Katrien Devreese

¹Coagulation Laboratory, Department of Clinical Chemistry, Microbiology and Immunology, Ghent University Hospital, Ghent, Belgium

,

Kathelijne Peerlinck

²Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium

,

Jef Arnout

²Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium

,

Marc F. Hoylaerts

²Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium

› Author Affiliations

Abstract

Summary

Lupus anticoagulants (LAC) consist of antiphospholipid antibodies, detected via their anticoagulant properties in vitro. Strong LAC relate to thromboembolic events, a hallmark of the anti-phospholipid syndrome. We have analyzed whether detection of this syndrome would benefit from thrombin generation measurements. Therefore, calibrated automated thrombography was done in normal plasma (n=30) and LAC patient plasma (n=48 non-anticoagulated, n=12 on oral anticoagulants), diluted 1:1 with a normal plasma pool. The anti-β₂-glycoprotein I monoclonal antibody 23H9, with known LAC properties, delayed the lag time and reduced the peak height during thrombin generation induction in normal plasma dose-dependently (0–150 μg/ml). At variance, LAC patient 1:1 plasma mixtures manifested variable lag time prolongations and/or peak height reductions. Coupling these two most informative thrombin generation parameters in a peak height/lag time ratio, and upon normalization versus the normal plasma pool, this ratio distributed normally and was reduced in the plasma mixtures, for 59/60 known LAC plasmas. The normalized peak height/lag time ratio correlated well with the normalized dilute prothrombin time, diluted Russell’s viper venom time and silica clotting time, measured in 1:1 plasma mixtures (correlation coefficients 0.59–0.72). The anticoagulant effects of activated protein C (0–7.5 nM) or 23H9 (0–150 μg/ml), spiked in the 1:1 LAC plasma mixtures were reduced for the majority of patients, compatible with functional competition between patient LAC and activated protein C and LAC and 23H9, respectively. Hence, the normalized thrombin generation-derived peak height/lag time ratio identifies LAC in plasma with high sensitivity in a single assay, irrespective of the patient’s treatment with oral anticoagulants.

Keywords

Thrombosis - α₂-glycoprotein I - lupus anticoagulants - prothrombin - antibodies

Full Text

References

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