Functionalized nanoparticles for nucleic acid sequence analysis using optical spectroscopies.
SERRS (surface-enhanced resonance Raman scattering) is a vibrational spectroscopy which allows extremely sensitive and selective detection of labelled DNA sequences with detection limits which rival, and in most cases surpass, that of fluorescence. SERRS relies on a visible chromophore adsorbing on to an enhancing surface. DNA itself is not SERRS-active, as it lacks a suitable visible chromophore and has poor adsorption properties on to the surfaces used for enhancement. The surface normally used for enhancement in these sorts of studies are metallic nanoparticles and, through modification of DNA probes by the addition of suitable SERRS labels, signals can be obtained that are highly sensitive and very selective. The aggregation state of the nanoparticles is critical to the sensitivity, and, in the present paper, we show how straightforward detection of labelled DNA probes can be achieved using SERRS in a quantitative manner and with a variety of different commercially available labels. In a second approach, we show how the properties of aggregation to turn on the SERRS effect can be exploited through DNA hybridization to give identification of a particular DNA sequence. This approach lends itself to closed-tube formats and is a promising way forward for molecular diagnostics using SERRS.
Graham
Duncan
D
Department of Pure and Applied Chemistry, Centre for Molecular Nanometrology, WestCHEM, University of Strathclyde, Glasgow, UK. duncan.graham@strath.ac.uk
Faulds
Karen
K
Thompson
David
D
Mackenzie
Fiona
F
Stokes
Robert
R
Macaskill
Alexandra
A
D004247:DNA / Q000032:analysis; D015342:DNA Probes; D053758:Nanoparticles / Q000737:chemistry*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013059:Spectrum Analysis, Raman / Q000379:methods*
D015342:DNA Probes; D004247:DNA