Isothermal discrimination of single-nucleotide polymorphisms via real-time kinetic desorption and label-free detection of DNA using silicon photonic microring resonator arrays

Anal Chem. 2011 Sep 1;83(17):6827-33. doi: 10.1021/ac201659p. Epub 2011 Aug 11.

Abstract

We report a sensitive, label-free method for detecting single-stranded DNA and discriminating between single nucleotide polymorphisms (SNPs) using arrays of silicon photonic microring resonators. In only a 10 min assay, DNA is detected at subpicomole levels with a dynamic range of 3 orders of magnitude. Following quantitation, sequence discrimination with single nucleotide resolution is achieved isothermally by monitoring the dissociation kinetics of the duplex in real-time using an array of SNP-specific capture probes. By leveraging the capabilities of the microring resonator platform, we successfully generate multiplexed arrays to quickly screen for the presence and identity of SNPs and show the robustness of this methodology by analyzing multiple target sequences of varying GC content. Furthermore, we show that this technique can be used to distinguish both homozygote and heterozygote alleles.

Publication types

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

MeSH terms

  • Alleles
  • DNA / analysis*
  • Kinetics
  • Nucleic Acid Hybridization / methods
  • Oligonucleotide Array Sequence Analysis / methods*
  • Photons
  • Polymorphism, Single Nucleotide*
  • Silicon / chemistry*
  • Temperature

Substances

  • DNA
  • Silicon