Digital quantification using amplified single-molecule detection

Nat Methods. 2006 Sep;3(9):725-7. doi: 10.1038/nmeth916.

Abstract

We describe a scheme for biomolecule enumeration by converting nanometer-scale specific molecular recognition events mediated by rolling-circle amplification to fluorescent micrometer-sized DNA molecules amenable to discrete optical detection. Our amplified single-molecule detection (SMD) approach preserves the discrete nature of the molecular population, allowing multiplex detection and highly precise quantification of molecules over a dynamic range of seven orders of magnitude. We apply the method for sensitive detection and quantification of the bacterial pathogen Vibrio cholerae.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics*
  • DNA, Circular / chemistry
  • DNA, Circular / genetics*
  • Molecular Probe Techniques
  • Nanotechnology
  • Sensitivity and Specificity
  • Spectrometry, Fluorescence / methods
  • Vibrio cholerae / genetics*

Substances

  • DNA, Bacterial
  • DNA, Circular