Reagent and kit for determining global coagulability and hemostatic potential

Abstract

A reagent and kit are disclosed for determining if a patient is hypercoagulable, hypocoagulable or normal. The test involves providing a test sample from the patient and initiating coagulation in the sample in the presence of an activator, which is added to the sample in an amount which will result in intrinsic tenase-dependent fibrin. Then the formation of the intrinsic tenase-dependent fibrin polymerization is monitored over time so as to derive a time-dependent profile, with the results of the fibrin polymerization monitoring determining whether the patient is hypercoagulable, normal or hypocoagulable. The coagulation activator is added in an amount that triggers a thrombin explosion that is dependent on the propagation phase and amplification pathways. In this way, a single assay can assess the hemostatic potential of a sample.

1 - 51 . (canceled)  
     
     
         52 . A kit for assessing the hemostatic potential of a test sample comprising: 
 a coagulation activator at a concentration of 11 picomolar or less, or a coagulation activator and instructions for diluting the coagulation activator to a concentration of 11 picomolar or less;    vesicles;    a metal divalent cation or a metal salt capable of dissociating into a metal divalent cation;    instructions for adding the coagulation activator, metal cation or metal salt, and vesicles to a test sample, and instructions for assessing the hemostatic potential of the test sample.    
     
     
         53 . The kit of  claim 52 , wherein the vesicles are a plurality of different phospholipids.  
     
     
         54 . The kit of  claim 52 , wherein the coagulation activator comprises tissue factor at a concentration of 11 picomolar or less.  
     
     
         55 . The kit of  claim 54 , further comprising a calcium cation or calcium salt that dissociates into a calcium cation.  
     
     
         56 . The kit of  claim 52 , wherein the kit is capable of indicating a sample to be any of hypocoagulable, normal and hypercoagulable, depending upon the condition of the patient from which the sample was taken.  
     
     
         57 . The kit of  claim 52 , wherein the kit is capable of indicating a patient, from which the sample was taken, to have any of thrombotic tendency, hemorraghic tendency, or stasis, depending on the patient.  
     
     
         58 - 68 . (canceled)  
     
     
         69 . The kit of  claim 52 , wherein said coagulation activator is present at a concentration of 8 picomolars or less.  
     
     
         70 . The kit of  claim 69 , wherein said coagulation activator is at a concentration of 6 picomolars or less.  
     
     
         71 . The kit of  claim 70 , wherein said coagulation activator is at a concentration of 3 picomolars or less.  
     
     
         72 . The kit of  claim 52 , further comprising phospholipids at a concentration of from 10 to 300 micromolar and an activator of an anticoagulant pathway and instruction for adding the activator to the test sample.  
     
     
         73 . The kit of  claim 72 , wherein the phospholipids are at a concentration of from 50 to 200 micromolar and the activator of the anticoagulant pathway is an activator of protein C.  
     
     
         74 . The kit of  claim 73 , wherein the protein C activator is thromobomodulin at a concentration of 30 nanomolar or less.  
     
     
         75 . The kit of  claim 74 , wherein the thrombomodulin is at a concentration of from 5 to 20 nanomolar.  
     
     
         76 - 77 . (canceled)  
     
     
         78 . The kit of  claim 74 , wherein the thrombomodulin is provided separately from the coagulation activator.  
     
     
         79 . The kit of  claim 78 , wherein the thrombomodulin is provided mixed with heparin, heparin sulphate or heparin-like molecules.  
     
     
         80 . The kit of  claim 52 , wherein the coagulation activator is provided mixed with vesicles, the metal ions or metal salt are provided separately from the coagulation activator, and an activator of an anticoagulation pathway is provided separately from the metal ions or salt, and separately from the coagulation activator.  
     
     
         81 . The kit of  claim 80 , wherein in a first container is provided tissue factor at a concentration of 11 picomolars or less mixed with vesicles which are phospholipids at a concentration of from 10 to 300 picomolar, in a second container is provided a metal salt in a concentration of from 5 to 50 mM, and in a third container is provided the coagulation activator which is thrombomodulin at a concentration of 30 nanomolar or less.  
     
     
         82 . The kit of  claim 52 , wherein a first container is provided having the coagulation activator which is tissue factor at a concentration of 11 picomolar or less mixed with vesicles which are phopholipids at a concentration of from 10 to 300 picomolar, a second container is provided having a metal salt in a concentration of from 5 to 50 micromolar, and a third container having the coagulation activator which is tissue factor at a concentration of 11 picomolar or less mixed with vesicles which are phospholipids at a concentration of from 10 to 300 picomolar and an activator of an anticoagulant pathway which is thrombomodulin at a concentration of 30 nanomolar or less.  
     
     
         83 - 96 . (canceled)  
     
     
         97 . The kit of  claim 52 , wherein the vesicles comprise platelets, cellular debris, phospholipid vesicles or platelet microparticles.  
     
     
         98 . The kit of  claim 52 , wherein the vesicles are phospholipid vesicles prepared by dilution, sonication, dialysis or extrusion.  
     
     
         99 - 100 . (canceled)