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Published VersionCC BY V4.0, Open Access
Journal article
Rapid, sensitive, and specific, low-resource molecular detection of Hendra virus
One Health, Vol.16, pp.1-7
2023
Abstract
Efficient and accurate diagnosis of Hendra virus (HeV), a biosafety level 4 (BSL 4) pathogen and zoonotic disease, is of primary importance for surveillance and outbreak control in the Australian equine industry. Sporadic HeV spillover events pose a serious public health concern and are predicted to expand geographically, aligning with the moving distribution of the main reservoir hosts, the flying foxes. Here we describe the development of a low resource rapid Hendra test. The test used a fast and simple sample processing protocol followed by reverse transcription isothermal recombinase polymerase amplification (RT RPA) combined with lateral flow detection (LFD) technology. Results were obtained in 30 min and required only a heating block, ice, and pipettes for liquid handling. The one step sample processing protocol inactivated HeV in 2 min, providing a simple protocol that could enable safe testing outside of a laboratory. Analytical sensitivity testing demonstrated a detection limit of 1000 copies/μL of synthetic HeV RNA, and analytical specificity testing indicated assays did not detect other pathogens. Gamma irradiated HeV spiked in viral transport medium was detected with high sensitivity, down to 10,000 TCID50/mL, the equivalent of 18 RNA copies per reaction. Collectively, our data suggests that our rapid Hendra test offers a potential first line screening on site alternative to gold standard RT PCR detection, which requires samples to be shipped to central containment laboratories, thermocyclers and labour intensive viral RNA purification, with testing time of approximately four hours. Our rapid Hendra test provided performance and speed without compromising sensitivity and specificity, and could become a promising more accessible tool for testing under resource limited conditions for the veterinary community and thoroughbred industry.
Details
- Title
- Rapid, sensitive, and specific, low-resource molecular detection of Hendra virus
- Authors
- N.M. Pollak (Corresponding Author) - University of the Sunshine Coast, Queensland, Centre for BioinnovationG.A. Marsh (Author) - CSIRO Health and BiosecurityM. Olsson (Author) - University of the Sunshine Coast, Queensland, School of Science, Technology and EngineeringD. McMillan (Corresponding Author) - University of the Sunshine Coast, Queensland, Centre for BioinnovationJ. Macdonald (Corresponding Author) - University of the Sunshine Coast, Queensland, Centre for Bioinnovation
- Publication details
- One Health, Vol.16, pp.1-7
- Publisher
- Elsevier BV
- Date published
- 2023
- DOI
- 10.1016/j.onehlt.2023.100504
- ISSN
- 2352-7714
- Copyright note
- © 2023 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
- Organisation Unit
- University of the Sunshine Coast, Queensland; School of Science, Technology and Engineering; Centre for Bioinnovation
- Language
- English
- Record Identifier
- 99706997602621
- Output Type
- Journal article
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