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

Nature Protocols volume 2, pages 2888–2895 (2007)Cite this article

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 ∼3 d, whereas the DNA sensing takes less than 2 h to complete.

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Figure 1: CDS strategy for DNA detection.
Figure 2: Schematic representations of correct electrode positioning.
Figure 3
Figure 4: Electrochemical behavior of DNA-modified gold electrodes.
Figure 5: DNA target concentration-dependent curves for the CDS.
Figure 6: Comparison of signal intensity for a series of targets associated with BRCA-1 at 10 pM: the perfectly matched DNA (A) and single-nucleotide mismatched DNA (T, C, G).
Figure 7: Comparison of both probe density and hybridization efficiency after three cycles of DNA hybridization and regeneration.

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Acknowledgements

This work was supported by National Natural Science Foundation (60537030 and 20725516), Shanghai Municipal Commission for Science and Technology (0652nm006 and 0752nm021), National Basic Research Program of China (2006CB933000 and 2007CB936000), Shanghai Rising-Star Program and Chinese Academy of Sciences.

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  1. Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai, 201800, China

    Jiong Zhang, Shiping Song, Lihua Wang, Dun Pan & Chunhai Fan

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Correspondence to Chunhai Fan.

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Zhang, J., Song, S., Wang, L. et al. A gold nanoparticle-based chronocoulometric DNA sensor for amplified detection of DNA. Nat Protoc 2, 2888–2895 (2007). https://doi.org/10.1038/nprot.2007.419

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