Field-deployable, Quantitative, Rapid Identification of Active Ebola Virus Infection in Unprocessed Blood

Shah, Kavit, Bentley, Emma, Tyler, Adam, Richards, Kevin, Wright, Edward, Easterbrook, Linda, Lee, Diane, Cleaver, Claire, Usher, Louise, Burton, Jane, Pitman, James, Bruce, Christine, Norwood, David and Moschos, Sterghios (2017) Field-deployable, Quantitative, Rapid Identification of Active Ebola Virus Infection in Unprocessed Blood. Chemical Science, 8. pp. 7780-7797. ISSN 2041-6539

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Official URL: http://dx.doi.org/10.1039/C7SC03281A

Abstract

The West African Ebola virus outbreak underlined the importance of delivering mass diagnostic capability outside the clinical or primary care setting in effectively containing public health emergencies caused by infectious disease. Yet, to date, there is no solution for reliably deploying at the point of need the gold standard diagnostic method, real time quantitative reverse transcription polymerase chain reaction (RT-qPCR), in a laboratory infrastructure-free manner. In this proof of principle work, we demonstrate direct performance of RT-qPCR on fresh blood using far-red fluorophores to resolve fluorogenic signal inhibition and controlled, rapid freeze/thawing to achieve viral genome extraction in a single reaction chamber assay. The resulting process is entirely free of manual or automated sample pre-processing, requires no microfluidics or magnetic/mechanical sample handling and thus utilizes low cost consumables. This enables a fast, laboratory infrastructure-free, minimal risk and simple standard operating procedure suited to frontline, field use. Developing this novel approach on recombinant bacteriophage and recombinant human immunodeficiency virus (HIV; lentivirus), we demonstrate clinical utility in symptomatic EBOV patient screening using live, infectious filoviruses and surrogate patient samples. Moreover, we evidence assay co-linearity independent of viral particle structure that may enable viral load quantification through pre-calibration, with no loss of specificity across an 8 log-linear maximum dynamic range. The resulting quantitative rapid identification (QuRapID) molecular diagnostic platform, openly accessible for assay development, meets the requirements of resource-limited countries and provides a fast response solution for mass public health screening against emerging biosecurity threats.

Item Type: Article
Uncontrolled Keywords: point of care, point of need, diagnostics, nucleic acid amplification testing, Ebola, Ebolavirus, West Africa, pseudovirus, public health england, united states army medical research institute for infectious diseases, screening, tool, ASSURED
Subjects: A300 Clinical Medicine
B800 Medical Technology
C500 Microbiology
C700 Molecular Biology, Biophysics and Biochemistry
H600 Electronic and Electrical Engineering
Department: Faculties > Health and Life Sciences > Applied Sciences
Depositing User: Sterghios Moschos
Date Deposited: 25 Sep 2017 11:40
Last Modified: 15 Nov 2017 09:48
URI: http://nrl.northumbria.ac.uk/id/eprint/31942

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