title: Label-free multiplexed virus detection using spectral reflectance imaging.
abstract: We demonstrate detection of whole viruses and viral proteins with a new label-free platform based on spectral reflectance imaging. The Interferometric Reflectance Imaging Sensor (IRIS) has been shown to be capable of sensitive protein and DNA detection in a real time and high-throughput format. Vesicular stomatitis virus (VSV) was used as the target for detection as it is well-characterized for protein composition and can be modified to express viral coat proteins from other dangerous, highly pathogenic agents for surrogate detection while remaining a biosafety level 2 agent. We demonstrate specific detection of intact VSV virions achieved with surface-immobilized antibodies acting as capture probes which is confirmed using fluorescence imaging. The limit of detection is confirmed down to 3.5 × 10(5)plaque-forming units/mL (PFUs/mL). To increase specificity in a clinical scenario, both the external glycoprotein and internal viral proteins were simultaneously detected with the same antibody arrays with detergent-disrupted purified VSV and infected cell lysate solutions. Our results show sensitive and specific virus detection with a simple surface chemistry and minimal sample preparation on a quantitative label-free interferometric platform.
authors.affiliation: Department of Electrical & Computer Engineering, Boston University, Boston, MA 02215, USA.
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Lopez
authors.forename: George G
authors.initials: GG
authors.lastname: Daaboul
authors.forename: Rahul S
authors.initials: RS
authors.lastname: Vedula
authors.forename: Emre
authors.initials: E
authors.lastname: Ozkumur
authors.forename: David A
authors.initials: DA
authors.lastname: Bergstein
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Geisbert
authors.forename: Helen E
authors.initials: HE
authors.lastname: Fawcett
authors.forename: Bennett B
authors.initials: BB
authors.lastname: Goldberg
authors.forename: John H
authors.initials: JH
authors.lastname: Connor
authors.forename: M Selim
authors.initials: MS
authors.lastname: Unlü
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D005453:Fluorescence; D007368:Interferometry / Q000379:methods*; D012680:Sensitivity and Specificity; D014721:Vesicular stomatitis Indiana virus / Q000302:isolation & purification*; D014764:Viral Proteins / Q000032:analysis; D014771:Virion / Q000302:isolation & purification
chemical_list: D014764:Viral Proteins