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United States Department of Agriculture

Agricultural Research Service

Research Project: COUNTERMEASURES TO CONTROL AND ERADICATE RIFT VALLEY FEVER (RFV)

Location: Arthropod-Borne Animal Diseases Research

Title: Surface-enhanced raman scattering detection of DNAs derived from virus genomes using au-coated paramagnetic nanoparticles

Authors
item Zhang, H -
item Harpster, Mark -
item WILSON, WILLIAM
item Johnson, Patrick -

Submitted to: Langmuir
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: January 25, 2012
Publication Date: January 25, 2012
Repository URL: http://pubs.acs.org/doi/abs/10.1021/la204890t
Citation: Zhang, H., Harpster, M.H., Wilson, W.C., Johnson, P.A. 2012. Surface-enhanced raman scattering detection of DNAs derived from virus genomes using au-coated paramagnetic nanoparticles. Langmuir. 28 (8):4030–4037.

Interpretive Summary: A model paramagnetic nanoparticle (MNP) duplex assay was demonstrated for West Nile virus (WNV) and Rift Valley fever virus genome that due to short reaction times can be conduced reproducibly and is amenable to adaptation within a portable, user-friendly detection platform for nucleic acids.

Technical Abstract: A magnetic capture-based, surface-enhanced Raman scattering (SERS) assay for DNA detection has been developed which utilizes Au-coated paramagnetic nanoparticles (Au@PMPs) as both a SERS substrate and effective bio-separation reagent for the selective removal of target DNAs from solution. Hybridization reactions contained two target DNAs, sequence complementary reporter probes conjugated with spectrally distinct Raman dyes distinct for each target, and Au@PMPs conjugated with sequence complementary capture probes. In this case, target DNAs were derived from the RNA genomes of the Rift Valley Fever virus (RVFV) or West Nile virus (WNV). The hybridization reactions were incubated for a short period and then concentrated within the focus beam of an interrogating laser by magnetic pull-down. The attendant SERS response of each individually captured DNA provided a limit of detection sensitivity in the range of 20-100 nM. X-ray diffraction and UV-vis analysis validated both the desired surface plasmon resonance properties and bi-metallic composition of synthesized Au@PMPs, and UV-vis spectroscopy confirmed conjugation of the Raman dye compounds malachite green (MG)and erythrosin B (EB) with the RVFV and WNV reporter probes, respectively. Finally, hybridization reactions assembled for multiplexed detection of both targets yielded mixed MG/EB spectra and clearly differentiated peaks which facilitate the quantitative detection of each DNA target. Based on the simple design of a single-particle DNA detection assay, the opportunity is provided to develop magnetic capture-based SERS assays that are easily assembled and adapted for high-level multiplex detection using low-cost Raman instrumentation.

Last Modified: 7/28/2014
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