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Journal article
Development and pre-clinical evaluation of a Zika virus diagnostic for low resource settings
Frontiers in Microbiology, Vol.14, pp.1-10
2023
Abstract
Zika virus (ZIKV) is a re-emerging flavivirus that poses a significant public health threat. ZIKV exhibits a wide array of non-vector borne human transmission routes, such as sexual transmission, transplacental transmission and blood transfusion. Detection and surveillance of ZIKV is considered paramount in prevention of major outbreaks. With the majority of cases reported in low-resource locations, simple, low-cost detection methods are considered highly desirable. Here we have developed a sensitive and specific ZIKV diagnostic using reverse transcription recombinase-aided amplification (RT-RAA) coupled with lateral flow detection (LFD) targeting a highly conserved region of the ZIKV NS1 gene. We show our rapid, isothermal-ZIKV-diagnostic (Iso-ZIKV-Dx) can detect 500 copies of synthetic ZIKV RNA/µL in under 30 minutes at a constant 39°C. Using mocksimulated urine samples, we observed that Iso-ZIKV-Dx also detects as low as 34.28 RNA copies/reaction of ZIKV (MR766 strain). Specificity testing confirmed that our test does not detect any co-circulating flaviviruses (dengue, West Nile, Japanese encephalitis, Murray Valley encephalitis and yellow fever viruses) or chikungunya virus. Furthermore, Ssample processing results show complete inactivation of ZIKV (MR766 strain) in 5 minutes at room temperature using our novel viral RNA sample preparation reagent. Furthermore, lateral flow strips testing demonstrates positive diagnoses in as little as 5 minutes in running buffer . Contrary to conventional RT-qPCR, our Iso-ZIKV-Dx does not require expensive machinery, specialised laboratory settings or extensively trained personnel. Our Pre-clinical evaluation demonstrates that our test exhibits robust, in-field capabilities without compromising sensitivity or specificity. When compared to the gold-standard RT-qPCR, our Iso-ZIKV-Dx test offers an array of applications that extend beyond diagnostics alone, including potential for surveillance and monitoring of ZIKV vector competency.
Details
- Title
- Development and pre-clinical evaluation of a Zika virus diagnostic for low resource settings
- Authors
- Rickyle Balea (Author) - University of the Sunshine Coast, Queensland, Centre for BioinnovationNina M Pollak (Corresponding Author) - University of the Sunshine Coast, Queensland, Centre for BioinnovationJody Hobson-Peters (Author) - The University of QueenslandJoanne Macdonald (Corresponding Author) - University of the Sunshine Coast, Queensland, Centre for BioinnovationDavid McMillan (Corresponding Author) - University of the Sunshine Coast, Queensland, Centre for Bioinnovation
- Publication details
- Frontiers in Microbiology, Vol.14, pp.1-10
- Publisher
- Frontiers Research Foundation
- Date published
- 2023
- DOI
- 10.3389/fmicb.2023.1214148
- ISSN
- 1664-302X
- Copyright note
- © 2023 Balea, Pollak, Hobson-Peters, Macdonald and Mcmillan. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
- Organisation Unit
- School of Health - Biomedicine; School of Science, Technology and Engineering; Centre for Bioinnovation; School of Health - Anatomy
- Language
- English
- Record Identifier
- 99976897802621
- Output Type
- Journal article
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