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Homogeneous detection of unamplified genomic DNA sequences based on colorimetric scatter of gold nanoparticle probes

Nature Biotechnology volume 22, pages 883–887 (2004)Cite this article

Abstract

Nucleic acid diagnostics is dominated by fluorescence-based assays that use complex and expensive enzyme-based target or signal-amplification procedures1,2,3,4,5,6. Many clinical diagnostic applications will require simpler, inexpensive assays that can be done in a screening mode. We have developed a 'spot-and-read' colorimetric detection method for identifying nucleic acid sequences based on the distance-dependent optical properties of gold nanoparticles. In this assay, nucleic acid targets are recognized by DNA-modified gold probes, which undergo a color change that is visually detectable when the solutions are spotted onto an illuminated glass waveguide. This scatter-based method enables detection of zeptomole quantities of nucleic acid targets without target or signal amplification when coupled to an improved hybridization method that facilitates probe-target binding in a homogeneous format. In comparison to a previously reported absorbance-based method7, this method increases detection sensitivity by over four orders of magnitude. We have applied this method to the rapid detection of mecA in methicillin-resistant Staphylococcus aureus genomic DNA samples.

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Figure 1: Colorimetric detection of nucleic acid sequences.
Figure 2: Comparison of detection methodologies for monitoring DNA-GNP binding to target DNA.
Figure 3: mecA gene detection with DNA-GNPs.
Figure 4: Colorimetric analysis using optical instrumentation.

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Acknowledgements

We thank the National Institutes of Health (2 R44 CA85008-02) for support of this work. We also thank Susan Hagenow for assistance in sequence design, and the manufacturing and engineering groups at Nanosphere for technical support.

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  1. Department of Applied Science, Nanosphere, Inc., 4088 Commercial Avenue, Northbrook, 60062, Illinois, USA

    James J Storhoff, Adam D Lucas, Viswanadham Garimella, Y Paul Bao & Uwe R Müller

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  1. James J Storhoff
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Correspondence to James J Storhoff.

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Nanosphere is a privately held company, and publication of the manuscript demonstrates scientific progress that could lead to future funding for the company.

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Storhoff, J., Lucas, A., Garimella, V. et al. Homogeneous detection of unamplified genomic DNA sequences based on colorimetric scatter of gold nanoparticle probes. Nat Biotechnol 22, 883–887 (2004). https://doi.org/10.1038/nbt977

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