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Nanomaterials 2015, 5(4), 1556-1570; doi:10.3390/nano5041556

A Paper-Based Sandwich Format Hybridization Assay for Unlabeled Nucleic Acid Detection Using Upconversion Nanoparticles as Energy Donors in Luminescence Resonance Energy Transfer

Chemical Sensors Group, Department of Chemical and Physical Sciences, University of Toronto Mississauga, 3359 Mississauga Road, Mississauga, ON L5L 1C6, Canada
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Author to whom correspondence should be addressed.
Academic Editor: Ming Su
Received: 24 July 2015 / Revised: 12 September 2015 / Accepted: 22 September 2015 / Published: 24 September 2015
(This article belongs to the Special Issue Nanomaterials for Biosensing Applications)
View Full-Text   |   Download PDF [3918 KB, uploaded 24 September 2015]   |  

Abstract

Bioassays based on cellulose paper substrates are gaining increasing popularity for the development of field portable and low-cost diagnostic applications. Herein, we report a paper-based nucleic acid hybridization assay using immobilized upconversion nanoparticles (UCNPs) as donors in luminescence resonance energy transfer (LRET). UCNPs with intense green emission served as donors with Cy3 dye as the acceptor. The avidin functionalized UCNPs were immobilized on cellulose paper and subsequently bioconjugated to biotinylated oligonucleotide probes. Introduction of unlabeled oligonucleotide targets resulted in a formation of probe-target duplexes. A subsequent hybridization of Cy3 labeled reporter with the remaining single stranded portion of target brought the Cy3 dye in close proximity to the UCNPs to trigger a LRET-sensitized emission from the acceptor dye. The hybridization assays provided a limit of detection (LOD) of 146.0 fmol and exhibited selectivity for one base pair mismatch discrimination. The assay was functional even in undiluted serum samples. This work embodies important progress in developing DNA hybridization assays on paper. Detection of unlabeled targets is achieved using UCNPs as LRET donors, with minimization of background signal from paper substrates owing to the implementation of low energy near-infrared (NIR) excitation. View Full-Text
Keywords: luminescence resonance energy transfer; upconversion nanoparticle; nucleic acid hybridization; sandwich assay; paper-based assay luminescence resonance energy transfer; upconversion nanoparticle; nucleic acid hybridization; sandwich assay; paper-based assay
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Zhou, F.; Noor, M.O.; Krull, U.J. A Paper-Based Sandwich Format Hybridization Assay for Unlabeled Nucleic Acid Detection Using Upconversion Nanoparticles as Energy Donors in Luminescence Resonance Energy Transfer. Nanomaterials 2015, 5, 1556-1570.

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