Single-quantum-dot-based DNA nanosensor.
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 kinetics. 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) 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 (approximately 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 mutation typical of some ovarian tumours in clinical samples.
Zhang
Chun-Yang
CY
Mechanical Engineering Department and Whitaker Biomedical Engineering Institute, The Johns Hopkins University, Baltimore, Maryland 21218, USA.
Yeh
Hsin-Chih
HC
Kuroki
Marcos T
MT
Wang
Tza-Huei
TH
D000818:Animals; D015374:Biosensing Techniques; D017931:DNA Primers / Q000737:chemistry; D004273:DNA, Neoplasm / Q000032:analysis*; D005260:Female; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry; D030342:Genetic Diseases, Inborn / Q000175:diagnosis; D006801:Humans; D036103:Nanotechnology; D010051:Ovarian Neoplasms / Q000175:diagnosis / Q000235:genetics*; D017354:Point Mutation; D045663:Quantum Dots; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004273:DNA, Neoplasm; D005456:Fluorescent Dyes