Journal article
Development and proof of concept evaluation for a low resource compatible Chikungunya virus diagnostic
PLoS Neglected Tropical Diseases, Vol.19(9), pp.1-15
2025
PMCID: PMC12448957
PMID: 40938967
Abstract
Chikungunya virus (CHIKV) is a positive sense RNA Alphavirus that continuous to pose major public health threats throughput the world. CHIKV is primarily transmitted via the Aedes genus mosquito, however, has also exhibited transmission routes via blood transfusion and vertical transmission (mother to child). With only one approved vaccine thus far and no approved medicines, early detection is crucial in mitigating CHIKV outbreaks. Here, we designed and evaluated a sensitive and specific CHIKV diagnostic using reverse transcription-recombinase aided amplification (RT-RAA) coupled lateral flow strip detection (LFD) targeting a highly conserved region of the CHIKV E1 gene. Our results demonstrate that using our simple sample preparation reagent (TNA-Cifer-E), can inactivate live CHIKV in two minutes at room temperature, whilst also sustaining viable viral RNA. Our specificity analysis demonstrates the Iso-CHIKV-Dx does not detect any closely related Alphaviruses nor any of the common co-circulating Flaviviruses. Proof-of-concept evaluation using urine spiked with CHIKV exhibited that in CHIKV infected urine samples, this Iso-CHIKV-Dx can detect as low as 570 copies/µL of CHIKV RNA in 30 minutes under isothermal conditions. Contrary to conventional RT-qPCR, our Iso-CHIKV-Dx does not require expensive machinery, advanced instrumentation or extensively trained personnel. Further performance comparisons also show that this Iso-CHIKV-Dx is four times faster than conventional RNA isolation and RT-qPCR. As such, pre-clinical evaluation demonstrates that this Iso-CHIKV-Dx has the potential to act as a robust, point of care CHIKV diagnostic that could prove to be highly beneficial in place of, or in the absence of RT-qPCR.
Details
- Title
- Development and proof of concept evaluation for a low resource compatible Chikungunya virus diagnostic
- Authors
- Rickyle Balea (Corresponding Author) - University of the Sunshine Coast, Queensland, School of Science, Technology and EngineeringAlberto A Amarilla - The University of QueenslandJody Hobson-Peters - The University of QueenslandJoanne Macdonald - University of the Sunshine Coast, Queensland, School of Science, Technology and EngineeringAndreas Suhrbier - QIMR Berghofer Medical Research InstituteVasilli M Kasimov - Griffith UniversityDaniel Watterson - The University of QueenslandNina M Pollak - University of the Sunshine Coast, Queensland, School of Science, Technology and EngineeringDavid J McMillan - University of the Sunshine Coast, Queensland, School of Science, Technology and Engineering
- Publication details
- PLoS Neglected Tropical Diseases, Vol.19(9), pp.1-15
- Publisher
- Public Library of Science
- Date published
- 2025
- DOI
- 10.1371/journal.pntd.0012352
- ISSN
- 1935-2735
- PMID
- 40938967; PMC12448957
- Copyright note
- © 2025 Balea et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
- Data Availability
- Data in this study can be freely obtained via download on ‘DataDryad.org’ DOI: https://doi.org/10.5061/dryad.547d7wmmf.
- Grants
- A single rapid field test for detecting Wolbachia, malaria and dengue in caught material, OPP1140133, Gates Foundation (United States, Seattle) - BMGF
- Grant note
- This study was supported by the Research Training Program (RTP) Commonwealth Research Scholarship (https://www.education.gov.au/research-block-grants/research-training-program), in accordance with the Higher Education Support act 2003 with the University of the Sunshine Coast (QLD, Australia) through R. B as the sole recipient.
- Organisation Unit
- School of Science, Technology and Engineering; Centre for Bioinnovation
- Language
- English
- Record Identifier
- 991162454602621
- Output Type
- Journal article
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