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Detection of rolling circle amplified DNA molecules using probe-tagged magnetic nanobeads in a portable AC susceptometer
Uppsala universitet, Teknisk-naturvetenskapliga vetenskapsområdet, Tekniska sektionen, Institutionen för teknikvetenskaper, Nanoteknologi och funktionella material.
Uppsala universitet, Science for Life Laboratory, SciLifeLab. Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Molekylära verktyg.
Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för immunologi, genetik och patologi, Molekylära verktyg. Uppsala universitet, Science for Life Laboratory, SciLifeLab.
Chalmers Industriteknik, Chalmers Science Park, Göteborg.
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2011 (Engelska)Ingår i: Biosensors & bioelectronics, ISSN 0956-5663, E-ISSN 1873-4235, Vol. 29, nr 1, s. 195-199Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Here, the volume-amplified magnetic nanobead detection assay (VAM-NDA) is for the first time applied for detection of rolling circle amplified (RCA) DNA molecules in a portable, commercial AC susceptometer that operates at ambient temperatures and with an analysis time of about 20 min. The performance of the assay is investigated using three different magnetic nanobead sizes: 50, 130 and 250 nm. The performance of the assay using the AC susceptometer is compared to the performance achieved using a superconducting quantum interference device (SQUID).

It is found that the performance of the assay is comparable in the two setups with a quantitative detection limit of ∼4 pM for all bead sizes under study.

The findings show that the VAM-NDA holds promise for future wide-spread implementation in commercial AC susceptometer setups thus opening up for the possibility to perform magnetic bead-based DNA detection in point-of-care and outpatient settings.

Ort, förlag, år, upplaga, sidor
2011. Vol. 29, nr 1, s. 195-199
Nyckelord [en]
Probe-tagged magnetic beads; Padlock probes; Rolling circle amplification; Brownian relaxation; AC susceptometer
Nationell ämneskategori
Nanoteknik Teknik och teknologier
Forskningsämne
Teknisk fysik med inriktning mot nanoteknologi och funktionella material; Teknisk fysik med inriktning mot fasta tillståndets fysik
Identifikatorer
URN: urn:nbn:se:uu:diva-159175DOI: 10.1016/j.bios.2011.08.019ISI: 000296174700031OAI: oai:DiVA.org:uu-159175DiVA, id: diva2:443095
Tillgänglig från: 2011-09-23 Skapad: 2011-09-23 Senast uppdaterad: 2017-12-08Bibliografiskt granskad
Ingår i avhandling
1. Detection of Biomolecules Using Volume-Amplified Magnetic Nanobeads
Öppna denna publikation i ny flik eller fönster >>Detection of Biomolecules Using Volume-Amplified Magnetic Nanobeads
2012 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Abstract [en]

This thesis describes a new approach to biomolecular analysis, called the volume-amplified magnetic nanobead detection assay (VAM-DNA). It is a sensitive, specific magnetic bioassay that offers a potential platform for the development of low-cost, easy-to-use diagnostic devices. The VAM-NDA consists of three basic steps: biomolecular target recognition, enzymatic amplification of the probe-target complex using the rolling circle amplification (RCA) technique, and addition of target complementary probe-tagged magnetic nanobeads which exhibit Brownian relaxation behavior. Target detection is demonstrated by measuring the frequency-dependent complex magnetization of the magnetic beads. The binding of the RCA products (target DNA-sequence coils) to the bead surface causes a dramatic increase in the bead size, corresponding essentially to the size of the DNA coil (typically around one micrometer). This causes a decrease in the Brownian relaxation frequency, since it is inversely proportional to the hydrodynamic size of the beads. The concentration of the DNA coils is monitored by measuring the decrease in amplitude of the Brownian relaxation peaks of free beads.

The parameters oligonucleotide surface coverage, bead concentration, bead size and RCA times were investigated in this thesis to characterize features of the assay. It was found that all of these parameters affect the outcome and efficiency of the assay.

The possibility of implementing the assay on a portable, highly sensitive AC susceptometer platform was also investigated. The performance of the assay under these circumstances was compared with that using a superconducting quantum interference device (SQUID); the sensitivity of the assay was similar for both platforms. It is concluded that, the VAM-NDA opens up the possibility to perform biomolecular detection in point-of-care and outpatient settings on portable platforms similar to the one tested in this thesis.

Finally, the VAM-NDA was used to detect Escherichia coli bacteria and the spores of Bacillus globigii, the non-pathogenic simulant of Bacillus anthracis. A limit of detection of at least 50 bacteria or spores was achieved. This shows that the assay has great potential for sensitive detection of biomolecules in both environmental and biomedical applications.

Ort, förlag, år, upplaga, sidor
Uppsala: Acta Universitatis Upsaliensis, 2012. s. 65
Serie
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 905
Nyckelord
Magnetic biosensor, magnetic nanobeads, Brownian relaxation, padlock probe, rolling circle amplification, DNA detection, protein detection
Nationell ämneskategori
Nanoteknik
Forskningsämne
Teknisk fysik med inriktning mot nanoteknologi och funktionella material
Identifikatorer
urn:nbn:se:uu:diva-169431 (URN)978-91-554-8288-6 (ISBN)
Disputation
2012-04-13, Å 2005, Ångströmlaboratoriet, Lägerhyddsvägen 1, Uppsala, 09:30 (Engelska)
Opponent
Handledare
Tillgänglig från: 2012-03-23 Skapad: 2012-02-29 Senast uppdaterad: 2012-03-29Bibliografiskt granskad

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Zardán Gómez de la Torre, TeresaMezger, AnjaHerthnek, DavidSvedlindh, PeterNilsson, MatsStrømme, Maria
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Nanoteknologi och funktionella materialScience for Life Laboratory, SciLifeLabMolekylära verktygFasta tillståndets fysik
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