title: Novel microfluidic platform for automated lab-on-chip testing of hypercoagulability panel.
abstract: Current methods for hypercoagulability panel testing require large blood volumes and long turn-around testing times. A novel microfluidic platform has been designed to perform automated multiplexed hypercoagulability panel testing at near patient, utilizing only a single droplet of blood sample. We test the hypothesis that this novel platform could be utilized to perform specific multiplexed ELISA-based hypercoagulability panel testing for antithrombin III, protein C, protein S and factor VIII antigens, as well as anticardiolipin/human anti-β2-glycoprotein-1 IgG antibodies--on blood samples. Sandwich ELISA was modified by utilizing magnetic beads coated with specific antibodies as the solid phase using fluorescence readout. Percentage recovery was calculated using four-parameter logistic curves. On-chip ELISA with single factors was compared with multiplex factor ELISA for known concentrations of sample. Blood samples were analyzed on-chip and compared with traditional bench-top assays. Time for multiplexed performance of hypercoagulability panel ELISA on-chip with controls is 72 min. Recovery rates (range 80-120%) for known concentrations of specific factors was not significantly different when assays were performed using a single factor vs. multiplex factor analysis. Assay results were not significantly different between individual assays performed either on bench-top or on-chip with patient blood and/or plasma. Utilizing a novel digital microfluidic platform, we demonstrate the feasibility of automated hypercoagulability panel testing on small volume of plasma and whole blood patient samples with high fidelity. Further investigation is required to test the application of this novel technology at point-of-care clinical settings.
authors.affiliation: Department of Cardiac Surgery, Children's Hospital Boston, Harvard Medical School, Boston, Massachusetts 02115, USA. sitaram.emani@cardio.chboston.org
authors.forename: Sirisha
authors.initials: S
authors.lastname: Emani
authors.forename: Ramakrishna
authors.initials: R
authors.lastname: Sista
authors.forename: Hugo
authors.initials: H
authors.lastname: Loyola
authors.forename: Cameron C
authors.initials: CC
authors.lastname: Trenor
authors.forename: Vamsee K
authors.initials: VK
authors.lastname: Pamula
authors.forename: Sitaram M
authors.initials: SM
authors.lastname: Emani
mesh_terms: D017153:Antibodies, Anticardiolipin / Q000097:blood; D000990:Antithrombin III / Q000032:analysis; D057205:Automation, Laboratory; D002138:Calibration; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation* / Q000379:methods; D005169:Factor VIII / Q000032:analysis; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D011486:Protein C / Q000032:analysis; D017293:Protein S / Q000032:analysis; D015203:Reproducibility of Results; D019851:Thrombophilia / Q000097:blood* / Q000175:diagnosis*; D053482:beta 2-Glycoprotein I / Q000097:blood / Q000276:immunology
chemical_list: D017153:Antibodies, Anticardiolipin; D007074:Immunoglobulin G; D011486:Protein C; D017293:Protein S; D053482:beta 2-Glycoprotein I; D000990:Antithrombin III; D005169:Factor VIII