Quantitative detection of antibiotic resistance genes using magnetic/luminescent core-shell nanoparticles

Ahjeong Son, Krassimira R. Hristova, Dosi Dosev, Ian M. Kennedy

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review

1 Scopus citations

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 DNA-NP 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 NP-hybridization were well correlated with the numbers measured by real-time qPCR assay (R2 = 0.92).

Original languageEnglish
Title of host publicationNanoscale Imaging, Sensing, and Actuation for Biomedical Applications V
DOIs
StatePublished - 2008
EventNanoscale Imaging, Sensing, and Actuation for Biomedical Applications V - San Jose, CA, United States
Duration: 21 Jan 200823 Jan 2008

Publication series

NameProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume6865
ISSN (Print)1605-7422

Conference

ConferenceNanoscale Imaging, Sensing, and Actuation for Biomedical Applications V
Country/TerritoryUnited States
CitySan Jose, CA
Period21/01/0823/01/08

Keywords

  • Antibiotic resistance
  • DNA
  • Hybridization-in-solution
  • Nanoparticles
  • Real-time qPCR
  • Tetracycline
  • Triclosan

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