A gold nanoparticle-based chronocoulometric DNA sensor for amplified detection of DNA

Nat Protoc. 2007;2(11):2888-95. doi: 10.1038/nprot.2007.419.

Abstract

We report a protocol for the amplified detection of target DNA by using a chronocoulometric DNA sensor (CDS). Electrochemistry is known to be rapid, sensitive and cost-effective; it thus offers a promising approach for DNA detection. Our CDS protocol is based on a 'sandwich' detection strategy, involving a capture probe DNA immobilized on a gold electrode and a reporter probe DNA loaded on gold nanoparticles (AuNPs). Each probe flanks one of two fragments of the target sequence. A single DNA hybridization event brings AuNPs, along with hundreds of reporter probes, in the proximity of the electrode. We then employ chronocoulometry to interrogate [Ru(NH3)6]3+ electrostatically bound to the captured DNA strands. This AuNP-amplified DNA sensor can selectively detect as low as femtomolar (zeptomoles) concentrations of DNA targets and conveniently analyze a breast cancer-associated BRCA-1 mutant DNA. The time range for the entire protocol is approximately 3 d, whereas the DNA sensing takes less than 2 h to complete.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques*
  • Chemistry Techniques, Analytical
  • DNA / analysis*
  • DNA / chemistry
  • DNA Mutational Analysis
  • Electrodes
  • Genes, BRCA1
  • Gold*
  • Humans
  • Hybridization, Genetic
  • Metal Nanoparticles*

Substances

  • Gold
  • DNA