Measuring thrombin activity in whole blood

摘要

The invention relates to a method for in vitro determining thrombin activity in a sample wherein the sample is a blood sample and thrombin generation is measured by the steps of: —contacting a layer of said sample with a fluorogenic substrate of thrombin, wherein said layer has a thickness within a range of 0.05 to 5 mm and a surface within a range of 10 to 500 mm2; —allowing thrombin to generate in said sample; —measuring the fluorescence emitted from the surface of the layer, by the fluorescent group released from the fluorogenic substrate as a result of enzymatic action of generated thrombin on said fluorogenic substrate.

主权项

1. A method for in vitro determining thrombin activity in a biological sample which is a sample of whole blood or a plasma sample and wherein thrombin generation is measured by the steps of:
contacting a layer of said sample with a fluorogenic substrate of thrombin, wherein said layer has a thickness within a range of about 0.05 to about 5 mm and a surface within a range of about 10 to 500 mm2 and wherein said sample is put into contact with a filter grid having a mesh size of about 50 to about 500 microns or packed spheres having a diameter of about 50 to about 500 microns which helps dispersion of the sample whereby sedimentation of the sample and retraction of the clot are decreased, allowing thrombin to generate in said sample, wherein said filter grid or packed spheres maintains the homogeneous dispersion of the sample after thrombin activation while reducing sedimentation of the sample and retraction of the clot within the assayed sample; and measuring the fluorescence emitted from the surface of the layer, by the fluorescent group released from the fluorogenic substrate as a result of enzymatic action of generated thrombin on said fluorogenic substrate.