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

JOSEPH TIBBS; BIN ZHAO; LAURA COOPER; YI LU; XIAOJING WANG; ANA SOL PEINETTI ; PRIYASH BARYA; XING WANG ; NANTAO LI; CONGNYU CHE; LEYANG LIU; LIJUN RONG; BRIAN T. CUNNINGHAM

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY

AMER CHEMICAL SOC

Lugar: Washington; Año: 2021

0002-7863

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.