Multiplexed efficient on-chip sample preparation and sensitive amplification-free detection of Ebola virus

K. Du, H. Cai, M. Park, T. A. Wall, M. A. Stott, K. J. Alfson, A. Griffiths, R. Carrion, J. L. Patterson, A. R. Hawkins, H. Schmidt, R. A. Mathies

    Research output: Contribution to journalArticlepeer-review

    26 Scopus citations

    Abstract

    An automated microfluidic sample preparation multiplexer (SPM) has been developed and evaluated for Ebola virus detection. Metered air bubbles controlled by microvalves are used to improve bead-solution mixing thereby enhancing the hybridization of the target Ebola virus RNA with capture probes bound to the beads. The method uses thermally stable 4-formyl benzamide functionalized (4FB) magnetic beads rather than streptavidin coated beads with a high density of capture probes to improve the target capture efficiency. Exploiting an on-chip concentration protocol in the SPM and the single molecule detection capability of the antiresonant reflecting optical waveguide (ARROW) biosensor chip, a detection limit of 0.021 pfu/mL for clinical samples is achieved without target amplification. This RNA target capture efficiency is two orders of magnitude higher than previous results using streptavidin beads and the limit of detection (LOD) improves 10×. The wide dynamic range of this technique covers the whole clinically applicable concentration range. In addition, the current sample preparation time is ~1 h which is eight times faster than previous work. This multiplexed, miniaturized sample preparation microdevice establishes a key technology that intended to develop next generation point-of-care (POC) detection system.

    Original languageEnglish (US)
    Pages (from-to)489-496
    Number of pages8
    JournalBiosensors and Bioelectronics
    Volume91
    DOIs
    StatePublished - May 15 2017

    Keywords

    • Air-bubble mixing
    • Microfluidics
    • Point-of-care
    • Single molecule RNA detection

    ASJC Scopus subject areas

    • Biotechnology
    • Biophysics
    • Biomedical Engineering
    • Electrochemistry

    Fingerprint Dive into the research topics of 'Multiplexed efficient on-chip sample preparation and sensitive amplification-free detection of Ebola virus'. Together they form a unique fingerprint.

    Cite this