A novel μ-fluidic whole blood coagulation assay based on Rayleigh surface-acoustic waves as a point-of-care method to detect anticoagulants.

A universal coagulation test that reliably detects prolonged coagulation time in patients, irrespective of the anticoagulant administered, has not been available to date. An easily miniaturised, novel μ-fluidic universal coagulation test employing surface acoustic waves (SAW) is presented here. SAW was employed to instantly mix and recalcify 6 μl citrated whole blood and image correlation analysis was used to quantify clot formation kinetics. The detection of clinically relevant anticoagulant dosing with old anticoagulants (unfractionated heparin, argatroban) and new anticoagulants (dabigatran, rivaroxaban) has been tested and compared to standard plasma coagulation assays. The applicability of this novel method has been confirmed in a small patient population. Coagulation was dose-proportionally prolonged with heparin, argatroban, dabigatran, and rivaroxaban, comparable to standard tests. Aspirin and clopidogrel did not interfere with the SAW-induced clotting time (SAW-CT), whereas the strong GPIIb/IIIa-inhibitor abciximab did interfere. Preliminary clinical data prove the suitability of the SAW-CT in patients being treated with warfarin, rivaroxaban, or dabigatran. The system principally allows assessment of whole blood coagulation in humans in a point-of-care setting. This method could be used in stroke units, emergency vehicles, general and intensive care wards, as well as for laboratory and home testing of coagulation.

[1]  Aydogan Ozcan,et al.  Imaging without lenses: achievements and remaining challenges of wide-field on-chip microscopy , 2012, Nature Methods.

[2]  M. Ray,et al.  A comparison of two APTT reagents which use silica activators. , 1989, Clinical and laboratory haematology.

[3]  Anthony J. Killard,et al.  Development of a fluorescent method for detecting the onset of coagulation in human plasma on microstructured lateral flow platforms. , 2011, The Analyst.

[4]  Leslie Y Yeo,et al.  A scaffold cell seeding method driven by surface acoustic waves. , 2007, Biomaterials.

[5]  W. Mueck,et al.  Safety, Tolerability, Pharmacodynamics, and Pharmacokinetics of Rivaroxaban—an Oral, Direct Factor Xa Inhibitor—Are Not Affected by Aspirin , 2006, Journal of clinical pharmacology.

[6]  Achim Wixforth,et al.  Microfluidic mixing via acoustically driven chaotic advection. , 2008, Physical review letters.

[7]  W. Mueck,et al.  Dose-escalation study of the pharmacokinetics and pharmacodynamics of rivaroxaban in healthy elderly subjects , 2008 .

[8]  H. Hemker,et al.  Pharmacokinetics and Pharmacodynamics of a Low Molecular Weight Heparin (Enoxaparin) after Subcutaneous Injection, Comparison with Unfractionated Heparin – A Three Way Cross Over Study in Human Volunteers , 1994, Thrombosis and Haemostasis.

[9]  K. Rathgen,et al.  The pharmacokinetics, pharmacodynamics and tolerability of dabigatran etexilate, a new oral direct thrombin inhibitor, in healthy male subjects. , 2007, British journal of clinical pharmacology.

[10]  K. Rathgen,et al.  Pharmacokinetics and Pharmacodynamics of Dabigatran Etexilate, an Oral Direct Thrombin Inhibitor, Are Not Affected by Moderate Hepatic Impairment , 2008, Journal of clinical pharmacology.

[11]  K. Rathgen,et al.  Pharmacokinetics and Pharmacodynamics of Dabigatran Etexilate, an Oral Direct Thrombin Inhibitor, With Coadministration of Digoxin , 2012, Journal of clinical pharmacology.

[12]  Christiane Ziegler,et al.  Investigation of prothrombin time in human whole-blood samples with a quartz crystal biosensor. , 2010, Analytical chemistry.

[13]  E. Favaloro,et al.  External Quality Assurance for Heparin Monitoring , 2012, Seminars in Thrombosis & Hemostasis.

[14]  E Theodorsson-Norheim,et al.  Kruskal-Wallis test: BASIC computer program to perform nonparametric one-way analysis of variance and multiple comparisons on ranks of several independent samples. , 1986, Computer methods and programs in biomedicine.

[15]  A. Hillarp,et al.  Considerations in the laboratory assessment of haemostasis , 2010, Haemophilia : the official journal of the World Federation of Hemophilia.

[16]  Gerhard Ziemer,et al.  Platelet aggregation monitoring with a newly developed quartz crystal microbalance system as an alternative to optical platelet aggregometry. , 2010, The Analyst.

[17]  I Lundström,et al.  Comparison of surface plasmon resonance and quartz crystal microbalance in the study of whole blood and plasma coagulation. , 2000, Biosensors & bioelectronics.

[18]  R. Ismagilov,et al.  Modular chemical mechanism predicts spatiotemporal dynamics of initiation in the complex network of hemostasis , 2006, Proceedings of the National Academy of Sciences.

[19]  G. Wensing,et al.  Safety, pharmacodynamics, and pharmacokinetics of single doses of BAY 59‐7939, an oral, direct factor Xa inhibitor , 2005, Clinical pharmacology and therapeutics.

[20]  S. Harder,et al.  Transition from argatroban to oral anticoagulation with phenprocoumon or acenocoumarol: effects on prothrombin time, activated partial thromboplastin time, and Ecarin Clotting Time , 2004, Thrombosis and Haemostasis.

[21]  Di Chen,et al.  A microfluidic chip for direct and rapid trapping of white blood cells from whole blood. , 2013, Biomicrofluidics.

[22]  H. Deckmyn,et al.  The CX3C chemokine fractalkine mediates platelet adhesion via the von Willebrand receptor glycoprotein Ib. , 2011, Blood.

[23]  Gary Chinga,et al.  Quantification of paper mass distributions within local picking areas , 2007 .

[24]  P. Renaud,et al.  Microfluidic System Based on Thermoexpandable Polymer for on Chip Blood Coagulation Testing , 2009 .

[25]  Fumihito Arai,et al.  Multiscale fabrication of a transparent circulation type blood vessel simulator. , 2010, Biomicrofluidics.

[26]  Leslie Y Yeo,et al.  Ultrafast microfluidics using surface acoustic waves. , 2009, Biomicrofluidics.

[27]  M. Ellmerer,et al.  Development and validation of a low cost blood filtration element separating plasma from undiluted whole blood. , 2012, Biomicrofluidics.

[28]  Kunihiro Hattori,et al.  In vitro evaluation of blood coagulation activation and microthrombus formation by a microchannel array flow analyzer. , 2004, Thrombosis research.

[29]  A. Zeiher,et al.  Anticoagulation With Argatroban for Elective Percutaneous Coronary Intervention: Population Pharmacokinetics and Pharmacokinetic‐Pharmacodynamic Relationship of Coagulation Parameters , 2011, Journal of clinical pharmacology.

[30]  Sascha Meyer dos Santos,et al.  Using ImageJ for the quantitative analysis of flow-based adhesion assays in real-time under physiologic flow conditions , 2010, Platelets.

[31]  A. Wixforth,et al.  Circulating but not immobilized N-deglycosylated von Willebrand factor increases platelet adhesion under flow conditions. , 2013, Biomicrofluidics.

[32]  Z. Ruggeri Von Willebrand factor: Looking back and looking forward , 2007, Thrombosis and Haemostasis.

[33]  Scott L Diamond,et al.  A membrane-based microfluidic device for controlling the flux of platelet agonists into flowing blood. , 2008, Lab on a chip.

[34]  W. Heerde,et al.  Global haemostasis assays, from bench to bedside. , 2012 .

[35]  A. Lombardi,et al.  Method for the determination of functional (clottable) fibrinogen by the new family of ACL coagulometers. , 1988, Thrombosis research.

[36]  Guillaume Mernier,et al.  Separation of platelets from other blood cells in continuous-flow by dielectrophoresis field-flow-fractionation. , 2011, Biomicrofluidics.

[37]  Rustem F Ismagilov,et al.  Microfluidics using spatially defined arrays of droplets in one, two, and three dimensions. , 2011, Annual review of analytical chemistry.

[38]  R. Paniccia,et al.  Point-of-Care Testing of Hemostasis in Cardiac Surgery , 2003, Thrombosis journal.

[39]  S. Hariharan,et al.  Clinical Pharmacology Basis of Deriving Dosing Recommendations for Dabigatran in Patients With Severe Renal Impairment , 2012, Journal of clinical pharmacology.

[40]  Yohsuke Imai,et al.  Asymmetry of red blood cell motions in a microchannel with a diverging and converging bifurcation. , 2011, Biomicrofluidics.

[41]  V. Ng Prothrombin time and partial thromboplastin time assay considerations. , 2009, Clinics in laboratory medicine.

[42]  A. Clemens,et al.  Pharmacology, Pharmacokinetics, and Pharmacodynamics of Dabigatran Etexilate, an Oral Direct Thrombin Inhibitor , 2009, Clinical and applied thrombosis/hemostasis : official journal of the International Academy of Clinical and Applied Thrombosis/Hemostasis.

[43]  A Alexander-Katz,et al.  Shear-induced unfolding triggers adhesion of von Willebrand factor fibers , 2007, Proceedings of the National Academy of Sciences.

[44]  D. Kubitza,et al.  Effects of the Oral, Direct Factor Xa Inhibitor Rivaroxaban on Platelet‐Induced Thrombin Generation and Prothrombinase Activity 1 , 2007, Journal of clinical pharmacology.

[45]  H. Weiss,et al.  Red blood cells: their dual role in thrombus formation. , 1980, Science.

[46]  W. Mueck,et al.  Body Weight Has Limited Influence on the Safety, Tolerability, Pharmacokinetics, or Pharmacodynamics of Rivaroxaban (BAY 59‐7939) in Healthy Subjects , 2007, Journal of clinical pharmacology.

[47]  Helen Song,et al.  On-chip titration of an anticoagulant argatroban and determination of the clotting time within whole blood or plasma using a plug-based microfluidic system. , 2006, Analytical chemistry.

[48]  I D Johnston,et al.  Whole blood pumping with a microthrottle pump. , 2010, Biomicrofluidics.

[49]  E. Kaliviotis,et al.  The effect of red blood cell aggregation on velocity and cell-depleted layer characteristics of blood in a bifurcating microchannel. , 2012, Biomicrofluidics.

[50]  J. Antaki,et al.  Investigation of platelet margination phenomena at elevated shear stress. , 2007, Biorheology.

[51]  W. Mueck,et al.  Safety, pharmacokinetics and pharmacodynamics of single/multiple doses of the oral, direct Factor Xa inhibitor rivaroxaban in healthy Chinese subjects. , 2009, British journal of clinical pharmacology.

[52]  A. Gamal,et al.  Miniaturized integration of a fluorescence microscope , 2011, Nature Methods.

[53]  Ji Jiang,et al.  Safety, pharmacokinetics and pharmacodynamics of single doses of rivaroxaban – an oral, direct factor Xa inhibitor – in elderly Chinese subjects , 2009, Thrombosis and Haemostasis.

[54]  Leslie Y Yeo,et al.  Surface acoustic wave concentration of particle and bioparticle suspensions , 2007, Biomedical microdevices.

[55]  T. Lindahl,et al.  Fluorescence-based blood coagulation assay device for measuring activated partial thromboplastin time. , 2011, Analytical chemistry.

[56]  Using microfluidics to understand the effect of spatial distribution of tissue factor on blood coagulation. , 2008, Thrombosis research.

[57]  J. Sadler,et al.  New concepts in von Willebrand disease. , 2005, Annual review of medicine.

[58]  A. Wixforth,et al.  Planar chip device for PCR and hybridization with surface acoustic wave pump. , 2005, Lab on a chip.

[59]  J. Sturm,et al.  Microfluidic device for label-free measurement of platelet activation. , 2008, Lab on a chip.