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Fluorescence-based blood coagulation assay device for measuring activated partial thromboplastin time
Magdalena M Dudek1, Nigel Kent, Kerstin M Gustafsson
1Biomedical Diagnostics Institute, National Centre for Sensor Research, Dublin City University, Dublin 9, Ireland.
Analytical Chemistry
|December 3, 2010
Summary
This study introduces a precise new method for measuring blood clotting time using a fluorescent probe and microstructured surfaces. The assay accurately detects fibrin clot formation, improving diagnostics for coagulation disorders and anticoagulant therapies.
Area of Science:
- Biomedical Engineering
- Clinical Chemistry
- Hematology
Background:
- Blood clotting time measurements are crucial for diagnosing coagulation disorders and managing anticoagulant therapies.
- Current clotting assays lack precision due to the variable nature of clot formation and sample matrices.
- Fibrin fiber formation is the key event in blood clot initiation.
Purpose of the Study:
- To develop a highly precise assay for detecting the onset of fibrin clot formation using a fluorescent probe.
- To apply this methodology to create an improved activated partial thromboplastin time (aPTT) test.
- To validate the assay's performance in monitoring anticoagulant effects, specifically heparin dosage.
Main Methods:
- Utilized a microstructured surface to enhance localized clot formation and fluorescent label redistribution.
- Employed a fluorescent probe for precise detection of fibrin clot onset in whole blood and plasma.
- Developed a lateral flow microfluidic platform for the activated partial thromboplastin time (aPTT) test.
Main Results:
- Demonstrated precise detection of fibrin clot formation onset.
- Achieved linearity (R(2)=0.996) for heparin dosage monitoring from 0 to 2 U/mL in spiked plasma.
- Showed high correlation with gold standard coagulometry (0.9986) and standard hospital aPTT (0.78 in 32 patient samples).
Conclusions:
- The developed fluorescent probe assay offers a precise and reliable method for measuring clotting time.
- The microfluidic aPTT test effectively monitors heparin dosage and correlates well with established methods.
- This technology has the potential to enhance the diagnosis and management of coagulation-related conditions.
