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 contribution

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 (US)
Title of host publicationProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume6865
DOIs
Publication statusPublished - 2008
EventNanoscale Imaging, Sensing, and Actuation for Biomedical Applications V - San Jose, CA, United States
Duration: Jan 21 2008Jan 23 2008

Other

OtherNanoscale Imaging, Sensing, and Actuation for Biomedical Applications V
CountryUnited States
CitySan Jose, CA
Period1/21/081/23/08

    Fingerprint

Keywords

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

ASJC Scopus subject areas

  • Engineering(all)

Cite this

Son, A., Hristova, K. R., Dosev, D., & Kennedy, I. M. (2008). Quantitative detection of antibiotic resistance genes using magnetic/luminescent core-shell nanoparticles. In Progress in Biomedical Optics and Imaging - Proceedings of SPIE (Vol. 6865). [68650P] https://doi.org/10.1117/12.764067