Zhang, Y, Pilapong, C, Guo, Y et al. (4 more authors) (2013) Sensitive, Simultaneous Quantitation of Two Unlabeled DNA Targets Using a Magnetic Nanoparticle-Enzyme Sandwich Assay. Analytical Chemistry, 85 (19). pp. 9238-9244. ISSN 0003-2700
Abstract
We report herein the development of a simple, sensitive colorimetric magnetic nanoparticle (MNP)- enzyme based DNA sandwich assay that is suitable for simultaneous label-free quantitation of two DNA targets down to 50 fM level. It can also effectively discriminate single-nucleotide polymorphisms (SNPs) in genes associated with human cancers (KRAS codon 12/13 SNPs). This assay uses a pair of specific DNA probes, one being covalently conjugated to a MNP for target capture while the other being linked to an enzyme for signal amplification, to sandwich a DNA target, allowing for convenient magnetic separation and subsequently efficient enzymatic signal amplification for high sensitivity. Careful optimization of the MNP surfaces and assay conditions greatly reduced the background, allowing for sensitive, specific detection of as little as 5 attomole (50 fM in 100 L) of target-DNA. Moreover, this sensor is robust, it can effectively discriminate cancer specific SNPs against the wild-type non-cancer target, and works efficiently in 10% human serum. Furthermore, this sensor can simultaneously quantitate two different DNA targets by using two pairs of unique capture- and signal- DNA probes specific for each target. This general, simple and sensitive DNA sensor appears to be well-suited for a wide range of genetics based biosensing and diagnostic applications.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2013 the authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
Keywords: | biosensing; magnetic nanoparticle; enzymatic assay; simultaneous DNA quantitation; single-nucleotide polymorphism |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) > School of Chemistry (Leeds) > Inorganic Chemistry (Leeds) The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) > School of Physics and Astronomy (Leeds) > Condensed Matter (Leeds) |
Funding Information: | Funder Grant number Leeds Cancer Research UK centre UNSPECIFIED The Wellcome Trust UNSPECIFIED Leeds Biomedical and Health Research Centre PF565 EPSRC EP/J501475/1 |
Depositing User: | Symplectic Publications |
Date Deposited: | 09 Sep 2013 09:48 |
Last Modified: | 11 Mar 2016 06:09 |
Published Version: | http://dx.doi.org/10.1021/ac402081u |
Status: | Published |
Publisher: | American Chemical Society |
Identification Number: | 10.1021/ac402081u |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:76322 |