Effectiveness of a new immuno-assay for the diagnosis of heparin-induced thrombocytopenia and improved specificity when detecting IgG antibodies

The diagnosis of heparin-induced thrombocytopenia (HIT) is based on clinical criteria and biological assays. Most immunoassays detect antibodies (either IgG alone or additionally IgA and IgM) against PF4 immobilised in wells of microtiter plates with stoichiometric concentrations of polyanion (heparin or polyvinylsulfonate). We studied whether diagnostic sensitivity and/or specificity for HIT could be improved using a novel assay in which unfractionated heparin is immobilised alone to the microwells, with PF4 (and, potentially, other heparin-dependent antigen proteins) provided by adding platelet lysate during the procedure. Samples from 101 patients with suspected HIT and from 101 controls (including 50 with antiphospholipid antibodies) were tested. The global assay (Zymutest HIA IgG/A/M (R), Hyphen BioMed) was positive for 39 of 40 patients with definite HIT (positive PF4-specific ELISA and positive serotonin release assay). It was positive in only two of the 101 control patients studied and also in 14 of the 61 patients with suspected HIT for whom the disease was excluded (specificity (sp): 77%). On the other hand, Zymutest HIA IgG (R), an IgG-specific assay, was positive in only six patients without HIT (Sp: 90%). Heparin-dependent IgG antibodies were present at higher levels in patients with definite HIT than in those for whom the diagnosis of HIT was ruled out. A single ELISA that detects IgG antibodies is more effective for the diagnosis of HIT in clinical practice. These results also support the hypothesis that heparin-dependent antibodies of IgG class have a major role in the pathogenesis of HIT.