Shortened 'Thrombin Time' Monitoring on QCM-D: A Better Substitute of 'Gold Standard'

Abstract

Thrombin is the central enzyme in the coagulation cascade. Recently, modern technologies for thrombin generation measurement have shown a multidisciplinary interest in methods and assays. This is first report to recognize and optimize for human plasma application of 'thrombin time' (TT) assay for quartz crystal microbalance with dissipation (QCM-D) technique. QCM-D technique has been studied comparatively with mechanical coagulometer (which is regarded as 'gold standard' for coagulation assays). The optimized protocol for TT has been applied for plasma samples (n=54) on both platforms. Thrombin times (TTs) on QCM-D platform versus those of 'gold standard' yielded a promising correlation line with R2 value of 0.86. For TT assay, QCM-D platform proved superior as compared to 'gold standard' platform due to following four edges. Firstly, QCM-D platform provides whole kinetic information, including monitoring of total coagulation on its measurement curve. 'Gold standard' cannot yield this information because it picks one point during coagulation. Secondly, TTs on QCM-D platform are 30% shorter as compared to those of 'gold standard'. Thirdly, TTs on QCM-D technique produced 16% lower %RSD demonstrating lower variability. Lastly, a historical lowest sample volume consumption of 2.50 µl has been applied on QCM-D platform. ‘Gold standard’ employs 40 times greater sample volume consumption for laboratory experiments of TT. Additionally, 40 times lower reagent volume consumption has been employed on QCM-D platform in comparison to its counterpart’s. These advantageous features are substantial support for point of care (POC) settings for TT assay via QCM-D technique.