Abstract 14693: Novel Microfluidic Platform for Point of Care Hypercoagulability Panel Testing
Introduction: Point-of-care diagnostic testing for hypercoagulability panel has potential to improve risk stratification and management of patients with cardiovascular diseases, but has been limited due to existing technology. A novel microfluidic platform constructed from printed circuit board technology has been designed to perform multiplexed hypercoagulability panel testing at near patient, utilizing only a single droplet of whole blood.
Hypothesis: We test the hypothesis that this novel platform could be utilized to perform specific ELISA based hypercoagulability assays — antithrombin III (ATIII), protein C, protein S, factor VIII, and anti-cardiolipin antibody — with only a few microliters of whole blood sample.
Methods: Whole blood sample and reagents were loaded onto the digital microfluidic platform, and all on-chip droplet manipulations were guided by custom designed software. Standard sandwich ELISA was modified by utilizing magnetic beads coated with antibodies specific to coagulation factor, biotin-labeled secondary antibody, streptavidin-alkaline phosphatase, Tris-tween 20 with 1mg/ml BSA wash buffer, and 4-methylumbelyferyl phosphate fluorescence substrate. Standard curves were developed, and on-chip assay performance was assessed using spiked whole blood samples, and recovery (%) was calculated utilizing 4-parameter logistic curves.
Results: Time for on-chip ELISA assay performance on 10 samples is 1hour 15 minutes compared to 6hrs for traditional bench top assay. Standard curves with whole blood recoveries for all tested factors range between 85% to 140% (Figure).
Conclusions: Utilizing a novel digital microfluidic platform, we demonstrate completely automated hypercoagulability panel testing on small volume of whole blood samples with high fidelity. Further investigation is required to test the application of this novel technology at point of care clinical settings.
- © 2010 by American Heart Association, Inc.