Digital-resolution detection of microRNA with singlebase selectivity by photonic resonator absorption microscopy

Taylor D. Canady, Nantao Li, Lucas D. Smith, Yi Lu, Manish Kohli, Andrew M. Smith, Brian T. Cunningham

Research output: Contribution to journalArticle

Abstract

Circulating exosomal microRNA (miR) represents a new class of blood-based biomarkers for cancer liquid biopsy. The detection of miR at a very low concentration and with single-base discrimination without the need for sophisticated equipment, large volumes, or elaborate sample processing is a challenge. To address this, we present an approach that is highly specific for a target miR sequence and has the ability to provide "digital" resolution of individual target molecules with high signal-to-noise ratio. Gold nanoparticle tags are prepared with thermodynamically optimized nucleic acid toehold probes that, when binding to a target miR sequence, displace a probe-protecting oligonucleotide and reveal a capture sequence that is used to selectively pull down the target-probe-nanoparticle complex to a photonic crystal (PC) biosensor surface. By matching the surface plasmon-resonant wavelength of the nanoparticle tag to the resonant wavelength of the PC nanostructure, the reflected light intensity from the PC is dramatically and locally quenched by the presence of each individual nanoparticle, enabling a form of biosensor microscopy that we call Photonic Resonator Absorption Microscopy (PRAM). Dynamic PRAM imaging of nanoparticle tag capture enables direct 100-aM limit of detection and single-base mismatch selectivity in a 2-h kinetic discrimination assay. The PRAM assay demonstrates that ultrasensitivity (<1 pM) and high selectivity can be achieved on a direct readout diagnostic.

Original languageEnglish (US)
Pages (from-to)19362-19367
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume116
Issue number39
DOIs
StatePublished - Sep 24 2019

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Optics and Photonics
MicroRNAs
Microscopy
Nanoparticles
Biosensing Techniques
Nucleic Acid Probes
Oligonucleotide Probes
Nanostructures
Signal-To-Noise Ratio
Tumor Biomarkers
Gold
Limit of Detection
Biopsy
Light
Equipment and Supplies

Keywords

  • Biosensors
  • Diagnostics
  • Liquid biopsy
  • Nanotechnology
  • Photonics

ASJC Scopus subject areas

  • General

Cite this

Digital-resolution detection of microRNA with singlebase selectivity by photonic resonator absorption microscopy. / Canady, Taylor D.; Li, Nantao; Smith, Lucas D.; Lu, Yi; Kohli, Manish; Smith, Andrew M.; Cunningham, Brian T.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 116, No. 39, 24.09.2019, p. 19362-19367.

Research output: Contribution to journalArticle

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