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Attomolar Zika virus oligonucleotide detection based on loop-mediated isothermal amplification and AC susceptometry
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Solid State Physics.
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Solid State Physics.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Clinical Immunology.
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Solid State Physics.
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2016 (English)In: Biosensors & bioelectronics, ISSN 0956-5663, E-ISSN 1873-4235, Vol. 86, p. 420-425Article in journal (Refereed) Published
Abstract [en]

Because of the serological cross-reactivity among the flaviviruses, molecular detection methods, such as reverse-transcription polymerase chain reaction (RT-PCR), play an important role in the recent Zika outbreak. However, due to the limited sensitivity, the detection window of RT-PCR for Zika viremia is only about one week after symptom onset. By combining loop-mediated isothermal amplification (LAMP) and AC susceptometry, we demonstrate a rapid and homogeneous detection system for the Zika virus oligonucleotide. Streptavidin-magnetic nanoparticles (streptavidin-MNPs) are premixed with LAMP reagents including the analyte and biotinylated primers, and their hydrodynamic volumes are dramatically increased after a successful LAMP reaction. Analyzed by a portable AC susceptometer, the changes of the hydrodynamic volume are probed as Brownian relaxation frequency shifts, which can be used to quantify the Zika virus oligonucleotide. The proposed detection system can recognize 1 aM synthetic Zika virus oligonucleotide in 20% serum with a total assay time of 27 min, which can hopefully widen the detection window for Zika viremia and is therefore promising in worldwide Zika fever control.

Place, publisher, year, edition, pages
2016. Vol. 86, p. 420-425
Keywords [en]
Zika virus, Loop-mediated isothermal amplification, Magnetic nanoparticles, Brownian relaxation, AC susceptometer
National Category
Biophysics Medical Biotechnology
Identifiers
URN: urn:nbn:se:uu:diva-307258DOI: 10.1016/j.bios.2016.06.085ISI: 000384853300057OAI: oai:DiVA.org:uu-307258DiVA, id: diva2:1046487
Funder
Swedish Research Council Formas, 221-2012-444 221-2014-574 2011-1692Available from: 2016-11-14 Created: 2016-11-11 Last updated: 2018-03-13Bibliographically approved
In thesis
1. Magnetic Nanoparticle Based Biosensors for Pathogen Detection and Cancer Diagnostics
Open this publication in new window or tab >>Magnetic Nanoparticle Based Biosensors for Pathogen Detection and Cancer Diagnostics
2018 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

This thesis describes several magnetic nanoparticle (MNP)-based biosensing strategies which take advantage of different magnetic sensors, molecular tools and nanotechnologies. Proposed biosensors can be classified into three groups, i.e., immunoassay-based, molecular amplification-based, and nanoparticle assembly-based. The principal motivation is to develop and optimize biosensors for out-of-lab and point-of-care testing.

Immunoassay-based biosensors described in this thesis employ antibodies as the bio-recognition element for the detection of bacteria cells/fragments or proteins. Two typical immunoassay formats, i.e., direct and competitive format, are studied and compared for bacteria detection. In addition, in the protein biomarker detection, MNP chains are formed in the presence of target analytes as well as in the external rotating magnetic field. The high shape/magnetic anisotropy of the chains provides better optomagnetic performance.

Two different molecular amplification methods, i.e., rolling circle amplification (RCA) and loop-mediated isothermal amplification (LAMP), are described under the topic of molecular amplification-based biosensors. In RCA-based biosensors, DNA probe modified MNPs bind to the amplicons after amplification. In LAMP-based biosensors, MNPs are either modified with primers that keep growing during the amplification, or are co-precipitated with the by-product (Mg2P2O7) of the amplification.

The design of the nanoparticle assembly-based biosensors described in this thesis is based on duplex-specific nuclease (DSN)-assisted target recycling and core-satellite magnetic superstructures. In the presence of target microRNA, DSN cuts the DNA scaffold of the core-satellite assembly, releasing MNP satellites that can be quantified by the sensor.

Different kinds of target analytes, i.e., pathogens or cancer biomarkers, are detected at the aiming for rapid, low-cost and user-friendly diagnostics.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2018. p. 55
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 1647
Keywords
Magnetic biosensors, magnetic nanoparticles, homogeneous assays, volumetric sensing
National Category
Other Engineering and Technologies not elsewhere specified
Research subject
Engineering Science with specialization in Solid State Physics
Identifiers
urn:nbn:se:uu:diva-346014 (URN)978-91-513-0278-2 (ISBN)
Public defence
2018-05-04, Häggsalen, Ångströmlaboratoriet, Lägerhyddsv. 1, Uppsala, 13:15 (English)
Opponent
Supervisors
Available from: 2018-04-13 Created: 2018-03-13 Last updated: 2018-04-24

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