title: Disposable cartridge platform for rapid detection of viral hemorrhagic fever viruses.
abstract: Light microscopy is a straightforward and highly portable imaging approach that is used for the detection of parasites, fungi, and bacteria. The detection of individual virus particles has historically not been possible through this approach. Thus, characterization of virus particles is typically performed using high-energy approaches such as electron microscopy. These approaches require purification of virions away from its normal milieu, significant levels of expertise, and only count a small number of particles at a time. To correct these deficiencies we created a platform that allows label-free, point-of-need virus imaging and counting. We adapted a multiplex-capable, interferometric imaging technique to a closed-system that allows real-time particle detection in complex mixtures. To maximize virus particle binding we constructed a disposable device with a constant flow rate of ∼3 μl min-1. Biosafety was achieved by having a sealable sample addition port. Using this platform we were able to readily identify virus binding in a 20 minute experiment. Sensitivity was comparable to laboratory-based assays such as ELISA and plaque assay, and showed equal or better sensitivity against paper-based assays designed for point-of-need use. Our results demonstrate a platform that can be used for rapid multiplexed detection and visualization of whole virus particles. We envision this technology as a sample-to-answer platform for detection and visualization of viruses without the need for prior labeling. This would enable both research investigation of virus particle behavior and morphology and have the potential to be used in a diagnostic context, where direct imaging from samples such as blood and urine would be valuable.
authors.affiliation: Department of Mechanical Engineering, Boston University, Boston, MA 02215, USA.
authors.forename: Steven M
authors.identifier: 0000-0001-7517-2112
authors.initials: SM
authors.lastname: Scherr
authors.affiliation: Nexgen Arrays, LLC, Boston, MA 02215, USA.
authors.forename: David S
authors.initials: DS
authors.lastname: Freedman
authors.affiliation: Galveston National Laboratory, University of Texas Medical Branch, Galveston, TX 77555, USA and Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555, USA.
authors.forename: Krystle N
authors.initials: KN
authors.lastname: Agans
authors.affiliation: Nexgen Arrays, LLC, Boston, MA 02215, USA.
authors.forename: Alexandru
authors.initials: A
authors.lastname: Rosca
authors.affiliation: Department of Microbiology and National Emerging Infectious Disease Laboratories, Boston University, Boston, MA 02118, USA. jhconnor@bu.edu.
authors.forename: Erik
authors.initials: E
authors.lastname: Carter
authors.affiliation: BD, Research Triangle Park, NC 27709, USA.
authors.forename: Melody
authors.initials: M
authors.lastname: Kuroda
authors.affiliation: Department of Mechanical Engineering, Boston University, Boston, MA 02215, USA.
authors.forename: Helen E
authors.initials: HE
authors.lastname: Fawcett
authors.affiliation: Galveston National Laboratory, University of Texas Medical Branch, Galveston, TX 77555, USA and Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555, USA.
authors.forename: Chad E
authors.initials: CE
authors.lastname: Mire
authors.affiliation: Galveston National Laboratory, University of Texas Medical Branch, Galveston, TX 77555, USA and Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555, USA.
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Geisbert
authors.affiliation: Department of Electrical Engineering, Boston University, Boston, MA 02215, USA and Department of Biomedical Engineering, Boston University, Boston, MA 02215, USA and Physics Department, Boston University, Boston, MA 02215, USA.
authors.forename: M Selim
authors.initials: MS
authors.lastname: Ünlü
authors.affiliation: Department of Microbiology and National Emerging Infectious Disease Laboratories, Boston University, Boston, MA 02118, USA. jhconnor@bu.edu and Department of Biomedical Engineering, Boston University, Boston, MA 02215, USA.
authors.forename: John H
authors.initials: JH
authors.lastname: Connor
mesh_terms: D029043:Ebolavirus / Q000302:isolation & purification*; D004867:Equipment Design; D007091:Image Processing, Computer-Assisted; D007368:Interferometry / Q000295:instrumentation*; D056656:Lab-On-A-Chip Devices; D057230:Limit of Detection; D008853:Microscopy; D010209:Paper; D015203:Reproducibility of Results; D014771:Virion / Q000302:isolation & purification; D014773:Virology / Q000295:instrumentation*