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Yoctomole electrochemical genosensing of Ebola virus cDNA by rolling circle and circle to circle amplification
Stockholm University, Faculty of Science, Department of Biochemistry and Biophysics. Stockholm University, Science for Life Laboratory (SciLifeLab).
Number of Authors: 42017 (English)In: Biosensors & bioelectronics, ISSN 0956-5663, E-ISSN 1873-4235, Vol. 93, p. 65-71Article in journal (Refereed) Published
Abstract [en]

This work addresses the design of an Ebola diagnostic test involving a simple, rapid, specific and highly sensitive procedure based on isothermal amplification on magnetic particles with electrochemical readout. Ebola padlock probes were designed to detect a specific L-gene sequence present in the five most common Ebola species. Ebola cDNA was amplified by rolling circle amplification (RCA) on magnetic particles. Further re-amplification was performed by circle-to-circle amplification (C2CA) and the products were detected in a double-tagging approach using a biotinylated capture probe for immobilization on magnetic particles and a readout probe for electrochemical detection by square-wave voltammetry on commercial screen-printed electrodes. The electrochemical genosensor was able to detect as low as 200 ymol, corresponding to 120 cDNA molecules of L-gene Ebola virus with a limit of detection of 33 cDNA molecules. The isothermal double-amplification procedure by C2CA combined with the electrochemical readout and the magnetic actuation enables the high sensitivity, resulting in a rapid, inexpensive, robust and user-friendly sensing strategy that offers a promising approach for the primary care in low resource settings, especially in less developed countries.

Place, publisher, year, edition, pages
2017. Vol. 93, p. 65-71
Keywords [en]
Ebola virus, Electrochemical genosensing, Magnetic particle, Isothermal amplification, Circle-to-circle amplification
National Category
Biological Sciences Environmental Biotechnology Chemical Sciences Nano Technology
Identifiers
URN: urn:nbn:se:su:diva-143424DOI: 10.1016/j.bios.2016.09.099ISI: 000399259000011PubMedID: 27838201OAI: oai:DiVA.org:su-143424DiVA, id: diva2:1107433
Conference
26th Anniversary World Congress on Biosensors, Gothenburg, Sweden, May 24-28, 2016
Available from: 2017-06-09 Created: 2017-06-09 Last updated: 2022-02-28Bibliographically approved

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Nilsson, Mats

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