期刊
ANALYTICAL BIOCHEMISTRY
卷 370, 期 2, 页码 186-194出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2007.08.001
关键词
DNA; hybridization-in-solution; nanoparticles; methyl tertiary-butyl ether; lanthanide oxide; magnetic particles
资金
- NIEHS NIH HHS [5 P42 ES04699-16] Funding Source: Medline
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:Gd-2-O-3 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. (c) 2007 Elsevier Inc. All rights reserved.
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