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Proceedings Paper

Quantitative detection of antibiotic resistance genes using magnetic/luminescent core-shell nanoparticles
Author(s): Ahjeong Son; Krassimira R. Hristova; Dosi Dosev; Ian M. Kennedy
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Paper Abstract

Nanoscale magnetic/luminescent core-shell particles were used for DNA quantification in a hybridization-in-solution format. We demonstrated a simple, high-throughput, and non-PCR based DNA assay for quantifying antibiotic resistance gene tetQ. Fe3O4/Eu:Gd2O3 nanoparticles (NPs) synthesized by spray pyrolysis were biofunctionalized by passive adsorption of NeutrAvidin. Following immobilization of biotinylated probe DNA on the particles' surfaces, target dsDNA and signaling probe DNA labeled with Cy3 were hybridized with NPs-probe DNA. Hybridized DNA complexes were separated from solution by a magnet, while non-hybridized DNA remained in solution. A linear quantification (R2 = 0.99) of a target tetQ gene was achieved based on the normalized fluorescence (Cy3/NPs) of DNANP hybrids. A real-time qPCR assay was used for evaluation of the NPs assay sensitivity and range of quantification. The quantity of antibiotic resistance tetQ genes in activated sludge microcosms, with and without addition of tetracycline or triclosan has been determined, indicating the potential of the optimized assay for monitoring the level of antibiotic resistance in environmental samples. In addition, the tetQ gene copy numbers in microcosms determined by NPhybridization were well correlated with the numbers measured by real-time qPCR assay (R2 = 0.92).

Paper Details

Date Published: 22 February 2008
PDF: 8 pages
Proc. SPIE 6865, Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications V, 68650P (22 February 2008); doi: 10.1117/12.764067
Show Author Affiliations
Ahjeong Son, Univ. of California, Davis (United States)
Krassimira R. Hristova, Univ. of California, Davis (United States)
Dosi Dosev, Univ. of California, Davis (United States)
Ian M. Kennedy, Univ. of California, Davis (United States)

Published in SPIE Proceedings Vol. 6865:
Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications V
Alexander N. Cartwright; Dan V. Nicolau, Editor(s)

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