Quantitative DNA hybridization in solution using magnetic/luminescent core-shell nanoparticles

Ahjeong Son, Dosi Dosev, Mikaela Nichkova, Zhiya Ma, Ian M. Kennedy, Kate M. Scow, Krassimira R. Hristova

Research output: Contribution to journalArticle

53 Scopus citations

Abstract

Nanoscale magnetic/luminescent core-shell particles were used for DNA quantification in a hybridization-in-solution approach. We demonstrated a rapid, simple, and non-polymerase chain reaction-based DNA hybridization-in-solution assay for quantifying bacteria capable of biodegrading methyl tertiary-butyl ether. Fe3O4/Eu:Gd2O3 core-shell nanoparticles synthesized by spray pyrolysis were biofunctionalized with NeutrAvidin. Following immobilization of a biotinylated probe DNA on the particles' surfaces via passive adsorption, target DNA labeled with fluorescein isothiocyanate was hybridized with probe DNA. The hybridized DNA complex was separated from solution with a magnet, while nonhybridized DNA remained in solution. The normalized fluorescence (fluorescein isothiocyanate/nanoparticles) measured with a spectrofluorometer indicated a linear quantification (R2 = 0.98) of the target bacterial 16 S rDNA. The rate of hybridization increased concurrently with the target DNA concentration. In addition, this approach differentiated between the signal outputs from perfectly complementary target and two-base mismatched target DNA in a range of concentrations, showing the specificity of the assay and the possibility for environmental applications.

Original languageEnglish (US)
Pages (from-to)186-194
Number of pages9
JournalAnalytical Biochemistry
Volume370
Issue number2
DOIs
StatePublished - Nov 15 2007

Keywords

  • DNA
  • Hybridization-in-solution
  • Lanthanide oxide
  • Magnetic particles
  • Methyl tertiary-butyl ether
  • Nanoparticles

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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  • Cite this

    Son, A., Dosev, D., Nichkova, M., Ma, Z., Kennedy, I. M., Scow, K. M., & Hristova, K. R. (2007). Quantitative DNA hybridization in solution using magnetic/luminescent core-shell nanoparticles. Analytical Biochemistry, 370(2), 186-194. https://doi.org/10.1016/j.ab.2007.08.001