Whole-blood thrombin generation monitored with a calibrated automated thrombogram-based assay

Area of Science:

• Hematology
• Biochemistry
• Clinical Diagnostics

Background:

• Calibrated automated thrombogram (CAT) assay in plasma is used for hypo- or hypercoagulable phenotypes.
• Current plasma-based assays do not reflect physiological conditions.
• Objective: Develop a whole blood CAT assay.

Purpose of the Study:

• To develop a reliable whole blood calibrated automated thrombogram (CAT) assay.
• To assess thrombin generation (TG) under more physiological conditions.
• To enable rapid screening of the hemostatic system.

Main Methods:

• Utilized thin-layer technology and a rhodamine 110-based thrombin substrate.
• Developed a whole blood CAT-based assay for thrombin generation measurement.
• Tested assay sensitivity, speed, and reliability.

Main Results:

• Whole blood CAT assay is sensitive and rapid for hemostatic system screening.
• Demonstrated low intra-assay coefficients of variation (CVs) for endogenous thrombin potential (6.7%) and thrombin peak height (6.5%).
• Erythrocytes, not platelets, significantly contribute to procoagulant membranes; significant correlations found in hemophilia A patients.

Conclusions:

• A reliable whole blood method for measuring thrombin generation (TG) has been developed.
• The assay requires only a drop of blood.
• Provides a useful measurement of TG under physiological conditions, superior to plasma assays.