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Resonance Energy Transfer-Based Nucleic Acid Hybridization Assays on Paper-Based Platforms Using Emissive Nanoparticles as Donors

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Biosensors and Biodetection

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1571))

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Abstract

Quantum dots (QDs) and upconverting nanoparticles (UCNPs) are luminescent nanoparticles (NPs) commonly used in bioassays and biosensors as resonance energy transfer (RET) donors. The narrow and tunable emissions of both QDs and UCNPs make them versatile RET donors that can be paired with a wide range of acceptors. Ratiometric signal processing that compares donor and acceptor emission in RET-based transduction offers improved precision, as it accounts for fluctuations in the absolute photoluminescence (PL) intensities of the donor and acceptor that can result from experimental and instrumental variations. Immobilizing NPs on a solid support avoids problems such as those that can arise with their aggregation in solution, and allows for facile layer-by-layer assembly of the interfacial chemistry. Paper is an attractive solid support for the development of point-of-care diagnostic assays given its ubiquity, low-cost, and intrinsic fluid transport by capillary action. Integration of nanomaterials with paper-based analytical devices (PADs) provides avenues to augment the analytical performance of PADs, given the unique optoelectronic properties of nanomaterials. Herein, we describe methodology for the development of PADs using QDs and UCNPs as RET donors for optical transduction of nucleic acid hybridization. Immobilization of green-emitting QDs (gQDs) on imidazole functionalized cellulose paper is described for use as RET donors with Cy3 molecular dye as acceptors for the detection of SMN1 gene fragment. We also describe the covalent immobilization of blue-emitting UCNPs on aldehyde modified cellulose paper for use as RET donors with orange-emitting QDs (oQDs) as acceptors for the detection of HPRT1 gene fragment. The data described herein is acquired using an epifluorescence microscope, and can also be collected using technology such as a typical electronic camera.

*These authors contributed equally to this work.

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Acknowledgments

The authors gratefully acknowledge the Natural Sciences and Engineering Research Council of Canada (NSERC) for financial support of their research. S.D. acknowledges NSERC for the provision of a graduate scholarship. M.O.N. is grateful to the Ontario Centres of Excellence (OCE) for provision of a Talent Edge postdoctoral fellowship. Y.H. acknowledges the Ministry of Training, Colleges and Universities for provision of an Ontario Graduate Scholarship (OGS).

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    Authors and Affiliations

    1. Chemical Sensors Group, Department of Chemical and Physical Sciences, University of Toronto Mississauga, 3359 Mississauga Road, Mississauga, ON, L5L 1C6, Canada

      Samer Doughan, M. Omair Noor, Yi Han & Ulrich J. Krull

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    1. Samer Doughan
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    2. M. Omair Noor
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    3. Yi Han
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    4. Ulrich J. Krull
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    Corresponding author

    Correspondence to Ulrich J. Krull .

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    Editors and Affiliations

    1. National Cancer Institute, National Institutes of Health, Rockville, Maryland, USA

      Avraham Rasooly

    2. National Cancer Institute, National Institutes of Health, Rockville, Maryland, USA

      Ben Prickril

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    Doughan, S., Noor, M.O., Han, Y., Krull, U.J. (2017). Resonance Energy Transfer-Based Nucleic Acid Hybridization Assays on Paper-Based Platforms Using Emissive Nanoparticles as Donors. In: Rasooly, A., Prickril, B. (eds) Biosensors and Biodetection. Methods in Molecular Biology, vol 1571. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6848-0_19

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    • DOI: https://doi.org/10.1007/978-1-4939-6848-0_19

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    • Publisher Name: Humana Press, New York, NY

    • Print ISBN: 978-1-4939-6846-6

    • Online ISBN: 978-1-4939-6848-0

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