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Optomagnetic bioassay for qualitative biplex detection of bacterial DNA sequences
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, Nanotechnology and Functional Materials.
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.
2015 (English)In: Biotechnology Journal, ISSN 1860-6768, E-ISSN 1860-7314, Vol. 10, no 3, 469-472 p.Article in journal (Refereed) Published
Abstract [en]

There is an increasing need to develop novel bioassay methods for low-cost, rapid, and easy-to-use multiplex detection of pathogens in various fields ranging from human infectious disease diagnosis, drinking water quality control, to food safety applications. Due to their unique advantages, magnetic and optomagnetic bioassay principles are particularly promising for biodetection platforms that will be used in developing countries. In this paper, an optomagnetic method for rapid and cost-efficient qualitative biplex detection of bacterial DNA sequences is demonstrated. Within less than two hours, the assay gives an answer to whether none, both, or only one of the bacterial DNA sequences is present in the sample. The assay relies on hybridization of oligonucleotide-functionalized magnetic nanobeads of two different sizes to rolling circle amplification (RCA) products originating from two different bacterial targets. The different bead sizes are equipped with different oligonucleotide probes, complementary to only one of the RCA products, and the read-out is carried out in the same sample volume. In an optomagnetic setup, the frequency modulation of transmitted laser light in response to an applied AC magnetic field is measured. The presented methodology is potentially interesting for low-cost screening of pathogens relating to both human and veterinary medicine in resource-poor regions of the world.

Place, publisher, year, edition, pages
2015. Vol. 10, no 3, 469-472 p.
National Category
Other Engineering and Technologies not elsewhere specified
Research subject
Engineering Science with specialization in Solid State Physics; Engineering Science with specialization in Nanotechnology and Functional Materials
Identifiers
URN: urn:nbn:se:uu:diva-233915DOI: 10.1002/biot.201400615ISI: 000351366600013PubMedID: 25512105OAI: oai:DiVA.org:uu-233915DiVA: diva2:754898
Funder
Swedish Research Council Formas, 221-2012-444Swedish Research Council Formas, 2011-1692Swedish Foundation for Strategic Research Carl Tryggers foundation , CTS 12:554
Available from: 2014-10-13 Created: 2014-10-13 Last updated: 2017-12-05Bibliographically approved
In thesis
1. Biomolecular Recognition Based on Field Induced Magnetic Bead Dynamics
Open this publication in new window or tab >>Biomolecular Recognition Based on Field Induced Magnetic Bead Dynamics
2014 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

In this thesis, three different read-out techniques for biomolecular recognition have been studied. All three techniques rely on the change in dynamic behaviour of probe functionalised magnetic beads after binding to a biomolecular target complementary to the probe.

In the first technique presented, the sample is exposed to an AC magnetic field while the response to this field is probed using a laser source and a photodetector positioned at opposite sides of the sample. Beads bound to the target entity will experience an increase in their hydrodynamic volume, and will not be able to respond as rapidly to an alternating field as free beads. Here, the target entity is either DNA coils formed by rolling circle amplification or biotinylated bovine serum albumin (bBSA). The change in dynamic behaviour is measured as a frequency dependent modulation of transmitted light. Limit of detections (LODs) of 5 pM DNA coils originating from a V. cholerae target and 100 pM of bBSA have been achieved.

In the second technique presented, the beads are magnetically transported across a probe functionalised detection area on a microchip. Beads bound to a target will be blocked from interaction with the detection area probes, whereas in the absence of a target, beads will be immobilised on the detection area. The LOD of biotin for this system proved to be in the range of 20 to 50 ng/ml.

In the third technique presented, the sample is microfluidically transported to a detection area on a microchip. The read-out is performed using a planar Hall effect bridge sensor. A sinusoidal current is applied to the bridge in one direction and the sensor output voltage is measured across the sensor in the perpendicular direction. The AC current induced bead magnetisation contributing to the sensor output will appear different for free beads compared to beads bound to a target. LODs of 500 B. globigii spores and 2 pM of V. cholerae DNA coils were achieved.

From a lab-on-a-chip point of view, all three techniques considered in this thesis show promising results with regards to sensitivity and integrability.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2014. 94 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 1193
Keyword
Magnetic biosensor, magnetic nanoparticle, DNA detection
National Category
Nano Technology Biochemistry and Molecular Biology Condensed Matter Physics
Research subject
Engineering Science
Identifiers
urn:nbn:se:uu:diva-234302 (URN)978-91-554-9077-5 (ISBN)
Public defence
2014-12-12, Häggsalen, Ångströmlaboratoriet, Lägerhyddsvägen 1, Uppsala, 09:15 (English)
Opponent
Supervisors
Funder
Knut and Alice Wallenberg FoundationSwedish Research Council
Available from: 2014-11-21 Created: 2014-10-15 Last updated: 2015-02-03

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