Heparin induced thrombocytopenia (HIT): platelet activation assay

A platelet aggregometry technique is employed as the functional assay for detecting HIT antibodies. Test serum is incubated together with freshly drawn normal donor platelets and either a high or low concentration of heparin. A patient with HIT antibodies will activate donor platelets in the presence of low concentration heparin but that effect is abolished at the higher concentration as the antibody is swamped.
Clinical details: 
Approximately 5% of patients on unfractionated heparin therapy develop type 2 heparin-induced thrombocytopenia (HIT). Some of the platelet factor 4 (PF4) released from activated platelets binds to the platelet surface, to which heparin will bind. This causes a conformational change in the PF4 and exposes neoepitopes which are immunogenic and can lead to antibody production. The thrombocytopenia arises from removal of antibody-coated platelets from the circulation by the reticuloendothelial system. Bleeding is rarely a problem yet conversely, thrombosis is a recognised complication because antibody binding activates platelets to form platelet aggregates, further reducing the platelet count. Procoagulant microparticles are generated and excess PF4 not bound to heparin instead binds to endothelial heparan sulphate which can lead to further antibody formation and immune complex-mediated endothelial damage, which can progress to thrombosis or DIC. Type 1 HIT is not immune mediated but caused by mild direct platelet activation by heparin and is ostensibly benign. HIT can occur in LMWH therapy but is less common.

HIT is largely a clinical diagnosis but laboratory assays are valuable for confirmation or exclusion. Immunological assays detect the antibodies directly whilst functional platelet activation assays demonstrate the effect of patient antibodies on donor platelets. Functional assays tend to have a lower sensitivity for HIT antibodies than immunological assays but a higher probability of identifying clinically significant antibodies.
Reference range: 


Sample type and Volume required: 
External requests: Citrated platelet poor plasma
400µL x 1 aliquot
Internal requests: please refer to EPR label
Turnaround time: 
7 - 10 days
Special sample instructions: 

The sample should be analysed or manipulated & stored in the laboratory within 4 hours of venepuncture. Please ensure sample tubes are filled exactly to the fill-line as underfilling creates a dilution error and leads to inaccurate results.

Diagnostic Haemostasis and Thrombosis Department
020 7188 2797
St Thomas' Hospital
North Wing - 4th and 5th Floors
Westminster Bridge Road
London SE1 7EH

Laboratory opening times
For clinical advice or interpretation of results, please contact the laboratory in the first instance.

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Last updated: 08/03/2017