Fluorescent silver nanocluster DNA probes for multiplexed detection using microfluidic capillary electrophoresis

Analyst. 2015 Mar 7;140(5):1609-15. doi: 10.1039/c4an01735h.

Abstract

DNA-stabilized fluorescent silver nanoclusters (AgNC DNA) are a new class of fluorophore that are formed by sequence specific interactions between silver and single-stranded DNA. By incorporating both target-binding and fluorescent-reporting sequences into a single synthetic DNA oligomer, AgNC DNA probes eliminate the need to conjugate dye or quencher molecules. In this study, we modify a AgNC DNA probe to demonstrate single-color multiplexed detection of DNA targets. We show that appending different lengths of poly-dT to the probe sequences tunes the electrophoretic mobility of AgNC DNA probes without affecting their fluorescence spectra. We use this to introduce a set of AgNC DNA probes selective for Hepatitis A, B and C target sequences that can be processed together in a simple, single-step protocol and distinguished with a resolution of 3.47 and signal to noise ratio of 17.23 in under 10 seconds by microfluidic capillary electrophoresis.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • DNA Probes / chemistry*
  • DNA, Single-Stranded / analysis
  • DNA, Single-Stranded / genetics
  • DNA, Viral / analysis*
  • DNA, Viral / genetics
  • Electrophoresis, Capillary / methods*
  • Fluorescence
  • Fluorescent Dyes / chemistry*
  • Fluorometry
  • Hepatitis / classification
  • Hepatitis / genetics
  • Hepatitis, Viral, Human / diagnosis
  • Hepatitis, Viral, Human / genetics
  • Hepatitis, Viral, Human / virology
  • Humans
  • Metal Nanoparticles / chemistry*
  • Microfluidics / methods*
  • Nanostructures / chemistry*
  • Polymerase Chain Reaction / methods
  • Silver / chemistry*

Substances

  • DNA Probes
  • DNA, Single-Stranded
  • DNA, Viral
  • Fluorescent Dyes
  • Silver