Label-Free Digital Detection of Intact Virions by Enhanced Scattering Microscopy

J Am Chem Soc. 2022 Feb 2;144(4):1498-1502. doi: 10.1021/jacs.1c09579. Epub 2021 Dec 20.

Abstract

Several applications in health diagnostics, food, safety, and environmental monitoring require rapid, simple, selective, and quantitatively accurate viral load monitoring. Here, we introduce the first label-free biosensing method that rapidly detects and quantifies intact virus in human saliva with single-virion resolution. Using pseudotype SARS-CoV-2 as a representative target, we immobilize aptamers with the ability to differentiate active from inactive virions on a photonic crystal, where the virions are captured through affinity with the spike protein displayed on the outer surface. Once captured, the intrinsic scattering of the virions is amplified and detected through interferometric imaging. Our approach analyzes the motion trajectory of each captured virion, enabling highly selective recognition against nontarget virions, while providing a limit of detection of 1 × 103 copies/mL at room temperature. The approach offers an alternative to enzymatic amplification assays for point-of-collection diagnostics.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Video-Audio Media

MeSH terms

  • Aptamers, Nucleotide / chemistry*
  • Biosensing Techniques / instrumentation
  • Biosensing Techniques / methods*
  • DNA / chemistry*
  • Humans
  • Immobilized Nucleic Acids / chemistry*
  • Limit of Detection
  • Microscopy / methods
  • Optics and Photonics / instrumentation
  • Optics and Photonics / methods
  • SARS-CoV-2 / chemistry
  • SARS-CoV-2 / isolation & purification*
  • Saliva / virology
  • Spike Glycoprotein, Coronavirus / chemistry

Substances

  • Aptamers, Nucleotide
  • Immobilized Nucleic Acids
  • Spike Glycoprotein, Coronavirus
  • spike protein, SARS-CoV-2
  • DNA