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Silica bead-based microfluidic device with integrated photodiodes for the rapid capture and detection of rolling circle amplification products in the femtomolar range
KTH, Centres, Science for Life Laboratory, SciLifeLab.
Stockholm Univ, Dept Biochem & Biophys, Sci Life Lab, SE-17165 Solna, Sweden..
Inst Engn Sistemas & Comp Microsistemas & Nanotec, Lisbon, Portugal.;IN Inst Nanosci & Nanotechnol, Lisbon, Portugal..
Stockholm Univ, Dept Biochem & Biophys, Sci Life Lab, SE-17165 Solna, Sweden..
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2019 (English)In: Biosensors & bioelectronics, ISSN 0956-5663, E-ISSN 1873-4235, Vol. 128, p. 68-75Article in journal (Refereed) Published
Abstract [en]

The rapid and sensitive detection of specific nucleic acid sequences at the point-of-care (PoC) is becoming increasingly in demand for a variety of emergent biomedical applications ranging from infectious disease diagnostics to the screening of antimicrobial resistance. To meet such demand, considerable efforts have been invested towards the development of portable and integrated analytical devices combining microfluidics with miniaturized signal transducers. Here, we demonstrate the combination of rolling circle amplification (RCA)-based nucleic acid amplification with an on-chip size-selective trapping of amplicons on silica beads (similar to 8 nL capture chamber) coupled with a thin-film photodiode (200 x 200 mu m area) fluorescence readout. Parameters such as the flow rate of the amplicon solution and trapping time were optimized as well as the photodiode measurement settings, providing minimum detection limits below 0.5 fM of targeted nucleic acids and requiring only 5 mu L of pre-amplified sample. Finally, we evaluated the analytical performance of our approach by benchmarking it against a commercial instrument for RCA product (RCP) quantification and further investigated the effect of the number of RCA cycles and elongation times (ranging from 10 to 120 min). Moreover, we provide a demonstration of the application for diagnostic purposes by detecting RNA from influenza and Ebola viruses, thus highlighting its suitability for integrated PoC systems.

Place, publisher, year, edition, pages
ELSEVIER ADVANCED TECHNOLOGY , 2019. Vol. 128, p. 68-75
Keywords [en]
Photodiodes, Rolling circle amplification, Infectious disease diagnostics, Microfluidics, Silica microbeads
National Category
Analytical Chemistry
Identifiers
URN: urn:nbn:se:kth:diva-244498DOI: 10.1016/j.bios.2018.12.004ISI: 000457950200010PubMedID: 30634076Scopus ID: 2-s2.0-85059537056OAI: oai:DiVA.org:kth-244498DiVA, id: diva2:1298035
Note

QC 20190321

Available from: 2019-03-21 Created: 2019-03-21 Last updated: 2019-04-04Bibliographically approved

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Soares, Ruben R. G.Russom, Aman

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