Evaluation of a flow cytometric assay for the confirmation of heparin-induced thrombocytopenia

Int J Lab Hematol. 2016 Jun;38(3):240-5. doi: 10.1111/ijlh.12475. Epub 2016 Mar 16.

Abstract

Introduction: Heparin-induced thrombocytopenia (HIT) is an immune-mediated thrombocytopenia associated with heparin therapy. The diagnosis consists of a combination of pretest probability and laboratory testing. The routinely available laboratory antigen binding assays for the detection of specific antibodies have a low HIT-positive predictive value; therefore, to exclude false-positive results, one of the functional assays should be performed. Functional assays evaluate the ability of heparin/PF4 antibodies to activate the platelets. The aim of our study was to validate the flow cytometric functional assay, based on the use of anti-CD61 and anti-CD62 antibodies, as a suitable diagnostic test for HIT.

Methods: Sera from patients with a clinical suspicion of HIT were previously analyzed with screening IgG-specific ELISA, and 41 of those which were positive were selected for the functional assay.

Results: Our results were compared to another functional assay - the HIPA (heparin-induced platelet aggregation assay). The diagnostic specificity of the flow cytometric assay was calculated based on HIPA results and was 83%.

Conclusion: Performing this functional test after the screening assay could significantly improve the specificity of HIT testing as heparin/PF4 antibodies are often not clinically significant.

Keywords: Heparin-induced thrombocytopenia; flow cytometry; functional assay; heparin-induced platelet aggregation assay; platelet activation.

Publication types

  • Clinical Trial

MeSH terms

  • Adult
  • Autoantibodies / blood
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Flow Cytometry / methods*
  • Flow Cytometry / standards
  • Heparin / adverse effects*
  • Humans
  • Immunoglobulin G / blood
  • Male
  • Platelet Aggregation*
  • Thrombocytopenia / blood*
  • Thrombocytopenia / chemically induced*

Substances

  • Autoantibodies
  • Immunoglobulin G
  • Heparin