A Simultaneous Monitoring of Coagulation Time and Fibrinogen via PiCT on QCM-D

Abstract

‘Prothrombinase induced Clotting Time’ (PiCT) has potential to detect all anticoagulants in clinics. In the present study, PiCT has been used as a tool for anticoagulant detection in human plasma on quartz crystal microbalance with dissipation (QCM-D) technique. QCM-D technique enables monitoring of PiCT point, total coagulation and fibrinogen concentration from frequency and dissipation curves in single set of measurements. This is impossible on mechanical coagulometer (which is considered as 'gold standard') technique, and it cannot yield coagulation and fibrinogen concentration from a single set of measurements. It requires additional Clauss method or modified Clauss method to calculate fibrinogen by employing different reagents and experimental setups. Additionally, the present report utilizes the lowest sample volume (and each reagent volume) consumptions of 1.66 µL. The sample/reagent volume consumption of 1.66 µL on QCM-D is 30 times lower in comparison with mechanical coagulometer’s that uses 50 µL for laboratory experiments for PiCT. This element is crucial for application of spot test via QCM-D in laboratory and clinics for Point of Care (POC) settings. Different doses of anticoagulant in 20 human plasma samples on QCM-D technique have been studied and compared in parallel to ‘gold standard’. PiCT on QCM-D technique yielded precise and accurate data. Additionally, both techniques produced % RSD values between 3 and 8.5 with slight fluctuations on both sides for PiCT points. The %RSD data for both techniques has lower variability for danaparoid. Furthermore, QCM-D technique enables monitoring of substantial fibrinogen concentrations (i.e. 1 - 6 g/L) with outstanding R2 value of 0.99 on the calibration curve. PiCT-QCM-D technique proved superior at all concentrations of fibrinogen in standard reference plasma for PiCT range (precision) on comparing to that of 'gold standard'.