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Direct Electrical Detection of sub-aM DNA concentrations
KTH, School of Electrical Engineering and Computer Science (EECS), Micro and Nanosystems.ORCID iD: 0000-0003-4364-794X
Science for Life Laboratory, Department of Biochemistry and Biophysics, Stockholm University, Stockholm.
Science for Life Laboratory, Department of Biochemistry and Biophysics, Stockholm University, Stockholm.
Science for Life Laboratory, Department of Biochemistry and Biophysics, Stockholm University, Stockholm.
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2019 (English)Article in journal (Other academic) Submitted
Place, publisher, year, edition, pages
2019.
National Category
Electrical Engineering, Electronic Engineering, Information Engineering Medical and Health Sciences Nano Technology
Identifiers
URN: urn:nbn:se:kth:diva-250773OAI: oai:DiVA.org:kth-250773DiVA, id: diva2:1313702
Note

QC 20190506

Available from: 2019-05-06 Created: 2019-05-06 Last updated: 2019-05-14Bibliographically approved
In thesis
1. Digital Electrical DNA Sensing
Open this publication in new window or tab >>Digital Electrical DNA Sensing
2019 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Molecule detection is a workhorse in life sciences and medicine, for example in cancer diagnosis and virus and bacterial detection. DNA analysis can provide vital information about the state of a host organism and its medical and health condition. A central challenge in DNA sensing lays in obtaining the following key detection characteristics in a single device: low limit of detection, small sample volume, high specificity, quantification, rapid time-to-result at a low cost.

Here we investigate whether direct electrical DNA sensing in a miniaturized detector can enable such performance. The detector consists of a gold-coated thin porous membrane, functionalized with oligonucleotides receptors, that is sandwiched between two off-stoichiometric thiol-ene-epoxy layers. The device works as follows. First, target DNA in the sample is specifically recognized by padlock probe hybridization and ligation. Second, the target-receptor circular molecules are amplified by rolling circle amplification (RCA), generating long ssDNA concatemers (RCP). Third, the RCPs are stretched through the membrane pores. Fourth, DNA metallization was used to form the gold nanowires bridging both sides of membrane pores after gold enhancement, which results in a conductive path that is measured with a simple resistance measurement. The thesis describes the engineering technology that enables low LoD detection of ssDNA using a digital measurement and details the development and optimization of the detector fabrication and operation, including structural design, materials, and microfluidic operation. We demonstrated a detector with sub-aM LoD, high specificity and simple operation in a miniaturized and uncomplicated format.

 Furthermore, the thesis studies the long-term liquid storage in nL scale well arrays fabricated in off-stoichiometric thiol-ene (OSTE). We demonstrated liquid storage with < 10 % loss of stored PBS buffer for 33 days and the on-demand electrically controlled liquid release.

The thesis presents the potential of a combination DNA detector with the method of liquid storage. Combining the on-chip liquid storage and DNA detection methods could provide a powerful alternative to conventional bio-detectors used in molecular diagnostics, and improved performance in multiplexed point-of-care sensing of (ultra-low abundant) biomolecules.

Place, publisher, year, edition, pages
Stockholm: KTH Royal Institute of Technology, 2019. p. 81
Series
TRITA-EECS-AVL ; 2019:38
Keywords
DNA detection, thiol-ene-epoxy, rolling circle amplification, digital measurement, DNA stretching, microwell-array, gold nanowires, electrical measurement, specificity, limit of detection, liquid storage, disease diagnosis, lab-on-a-chip, point-of-care.
National Category
Engineering and Technology Medical and Health Sciences
Identifiers
urn:nbn:se:kth:diva-250783 (URN)978-91-7873-162-6 (ISBN)
Public defence
2019-06-11, Q2, Malvinas väg 10, Stockholm, 10:00 (English)
Opponent
Supervisors
Note

QC 20190508

Available from: 2019-05-08 Created: 2019-05-06 Last updated: 2019-05-14Bibliographically approved

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Guo, Maoxiangvan der Wijngaart, Wouter

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