Oligonucleotide and polymer functionalized nanoparticles for amplification-free detection of DNA

Thomson, David A. C., Tee, Ernest H. L., Tran, Nguyen T. D., Monteiro, Michael J. and Cooper, Matthew A. (2012) Oligonucleotide and polymer functionalized nanoparticles for amplification-free detection of DNA. Biomacromolecules, 13 6: 1981-1989. doi:10.1021/bm300717f


Author Thomson, David A. C.
Tee, Ernest H. L.
Tran, Nguyen T. D.
Monteiro, Michael J.
Cooper, Matthew A.
Title Oligonucleotide and polymer functionalized nanoparticles for amplification-free detection of DNA
Journal name Biomacromolecules   Check publisher's open access policy
ISSN 1525-7797
1526-4602
Publication date 2012-06-01
Sub-type Article (original research)
DOI 10.1021/bm300717f
Open Access Status Not Open Access
Volume 13
Issue 6
Start page 1981
End page 1989
Total pages 9
Place of publication Washington, DC, United States
Publisher American Chemical Society
Language eng
Formatted abstract
Sensitive and quantitative nucleic acid testing from complex biological samples is now an important component of clinical diagnostics. Whereas nucleic acid amplification represents the gold standard, its utility in resource-limited and point-of-care settings can be problematic due to assay interferants, assay time, engineering constraints, and costs associated with both wetware and hardware. In contrast, amplification-free nucleic acid testing can circumvent these limitations by enabling direct target hybridization within complex sample matrices. In this work, we grew random copolymer brushes from the surface of silica-coated magnetic nanoparticles using azide-modified and hydroxyl oligo ethylene glycol methacrylate (OEGMA) monomers. The azide-functionalized polymer brush was first conjugated, via copper-catalyzed azide/alkyne cycloaddition (CuAAC), with herpes simplex virus (HSV)-specific oligonucleotides and then with alkyne-substituted polyethylene glycol to eliminate all residual azide groups. Our methodology enabled control over brush thickness and probe density and enabled multiple consecutive coupling reactions on the particle grafted brush. Brush- and probe-modified particles were then combined in a 20 min hybridization with fluorescent polystyrene nanoparticles modified with HSV-specific reporter probes. Following magnetic capture and washing, the particles were analyzed with an aggregate fluorescence measurement, which yielded a limit of detection of 6 pM in buffer and 60 pM in 50% fetal bovine serum. Adoption of brush- and probe-modified particles into a particle counting assay will result in the development of diagnostic assays with significant improvements in sensitivity.
Keyword Transfer radical polymerization
Glycol methacrylate
Paramagnetic nanoparticles
Ambient-temperature
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

 
Versions
Version Filter Type
Citation counts: TR Web of Science Citation Count  Cited 24 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 19 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Mon, 09 Jul 2012, 20:19:13 EST by System User on behalf of Institute for Molecular Bioscience