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Real time PCR is the mainstay of current nucleic acid assays, underpinning applications in forensic science, point-of-care diagnostics and detection of bioterrorism agents. Despite its broad utility, the search for new tests continues, inspired by second and third generation DNA sequencing technologies and fuelled by progress in single molecule fluorescence spectroscopy, nanotechnology and microfabrication. These new methods promise the direct detection of nucleic acids without the need for enzymatic amplification. In this feature article, we provide a chemist's perspective on this multidisciplinary area, introducing the concepts of single molecule detection then focussing on the selection of labels and probe chemistry suitable for generating a signal detectable by ultrasensitive fluorescence spectroscopy. Finally, we discuss the further developments that are required to incorporate these detection platforms into integrated 'sample-in-answer-out' instruments, capable of detecting many target sequences in a matter of minutes.

Original publication

DOI

10.1039/c0cc04215c

Type

Journal article

Journal

Chem Commun (Camb)

Publication Date

07/04/2011

Volume

47

Pages

3717 - 3735

Keywords

Animals, Equipment Design, Fluorescence Resonance Energy Transfer, Fluorescent Dyes, Humans, Microfluidic Analytical Techniques, Nucleic Acids, Sensitivity and Specificity, Sequence Analysis, DNA, Spectrometry, Fluorescence