Hemostasis assay

摘要

Provided is a hemostasis assay comprising a reaction mixture comprising a blood product to be tested, a trigger molecule for inducing thrombin generation, a thrombin-specific substrate which, upon cleavage by thrombin, produces a measurable thrombin-specific signal, a trigger molecule for inducing plasmin generation, a plasmin-specific substrate which, upon cleavage by plasmin, produces a measurable plasmin-specific signal, a phospholipid-containing surface, and calcium ions. The assay allows determination of the amount of thrombin and the amount of plasmin generated in the reaction mixture in time, starting at t=0, by measuring the thrombin-specific and plasmin-specific signals.

主权项

1. An in vitro method for determining thrombin potential and plasmin potential generated in a single enzymatic reaction mixture in a single well, the method comprising:
adding (i) a trigger molecule for inducing thrombin generation; (ii) a thrombin-specific substrate, which, upon cleavage by thrombin, produces a measurable, thrombin-specific signal; (iii) a trigger molecule for inducing plasmin generation; (iv) a plasmin-specific substrate, which, upon cleavage by plasmin, produces a measurable, plasmin-specific signal; (v) a phospholipid-containing surface; and (vi) calcium ions; to a test sample comprising a blood product, thereby generating the single enzymatic reaction mixture; simultaneously measuring thrombin-specific and plasmin-specific signals in the single enzymatic reaction mixture in the single well; and determining the thrombin potential and the plasmin potential generated in the single enzymatic reaction mixture in the single well.