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Single-quantum-dot-based DNA nanosensor

Abstract

Rapid and highly sensitive detection of DNA is critical in diagnosing genetic diseases. Conventional approaches often rely on cumbersome, semi-quantitative amplification of target DNA to improve detection sensitivity. In addition, most DNA detection systems (microarrays, for example), regardless of their need for target amplification, require separation of unhybridized DNA strands from hybridized stands immobilized on a solid substrate, and are thereby complicated by solution–surface binding kinetics1,2. Here, we report an ultrasensitive nanosensor based on fluorescence resonance energy transfer (FRET) capable of detecting low concentrations of DNA in a separation-free format. This system uses quantum dots (QDs)3,4,5 linked to DNA probes to capture DNA targets. The target strand binds to a dye-labelled reporter strand thus forming a FRET donor–acceptor ensemble. The QD also functions as a concentrator that amplifies the target signal by confining several targets in a nanoscale domain. Unbound nanosensors produce near-zero background fluorescence, but on binding to even a small amount of target DNA (50 copies or less) they generate a very distinct FRET signal. A nanosensor-based oligonucleotide ligation assay has been demonstrated to successfully detect a point mutation6 typical of some ovarian tumours in clinical samples.

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Figure 1: Schematic of single-QD-based DNA nanosensors.
Figure 2: Detection of single-dot FRET signals.
Figure 3: Characterization of FRET.
Figure 4: Evaluation of nanosensor performance.

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Acknowledgements

The authors thank Y. Peng, S. Yang, S. Lin and Y. P. Ho for valuable discussions, I. M. Shih for providing us with PCR products from clinical samples for Kras point mutation detections, and L. Brand and D. Toptygin for providing support with the TCSPC fluorescence lifetime measurements. This work was supported primarily by NSF under award no. DBI-0352407 and also by the Whitaker Foundation.

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Correspondence to Tza-Huei Wang.

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Zhang, CY., Yeh, HC., Kuroki, M. et al. Single-quantum-dot-based DNA nanosensor. Nature Mater 4, 826–831 (2005). https://doi.org/10.1038/nmat1508

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