Quantitative DNA hybridization in solution using magnetic/luminescent core-shell nanoparticles.
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 (R(2)=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.
Son
Ahjeong
A
Department of Land, Air, and Water Resources, University of California Davis, One Shields Avenue, Davis, CA 95616, USA.
Dosev
Dosi
D
Nichkova
Mikaela
M
Ma
Zhiya
Z
Kennedy
Ian M
IM
Scow
Kate M
KM
Hristova
Krassimira R
KR
D020137:Base Pair Mismatch; D001483:Base Sequence; D004247:DNA / Q000032:analysis* / Q000235:genetics*; D007700:Kinetics; D049449:Luminescence; D008280:Magnetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D053758:Nanoparticles; D009693:Nucleic Acid Hybridization; D012996:Solutions
D012996:Solutions; D004247:DNA