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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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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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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DOI: https://doi.org/10.1038/nbt977
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