Method of detecting platelet activating antibodies that cause heparin-induced thrombocytopenia/thrombosis
The present invention provides a method of detecting platelet activation in a patient, the method comprising the steps of a) obtaining a blood sample from a patient suspected of having heparin-induced thrombocytopenia (HIT); b) incubating an effective amount of platelet factor 4 (PF4) with a sample of platelets to yield a sample of PF4-treated platelets; c) contacting the patient blood sample with the PF4-treated platelets; and d) measuring the extent of platelet activation, wherein an increase in platelet activation compared with results obtained using a normal blood sample is indicative of the patient having HIT.
1. A kit for determining the presence of platelet-activating pathogenic heparin-induced thrombocytopenia (HIT) antibodies as opposed to non-activating non-pathogenic HIT antibodies in a whole blood, plasma or serum sample from a patient suspected of having HIT comprising:
a) purified platelet factor 4 (PF4);
b) an antibody or probe capable of detecting platelet activation or antibody bound to platelets;
c) instructions for use;
d) an effective amount of whole blood, plasma, or serum control sample negative for platelet-activating pathogenic HIT antibodies; and
e) an effective amount of whole blood, plasma, or serum control sample positive for platelet-activating pathogenic HIT antibodies.
2. The kit of claim 1 further comprising at least 50 U/ml of heparin.
3. The kit of claim 1 further comprising an inhibitor of platelet activation.
4. The kit of claim 3 wherein the inhibitor of platelet activation is prostaglandin E1.
5. The kit of claim 1 wherein the antibody or probe is fluorescently labeled.
6. The kit of claim 5 wherein the antibody or probe is fluorescent-labeled annexin V.
7. The kit of claim 5 wherein the antibody or probe is fluorescent-labeled anti-p-selectin antibody.