Development and proof of concept evaluation for a low resource compatible Chikungunya virus diagnostic

Rickyle Balea; Alberto A Amarilla; Jody Hobson-Peters; Joanne Macdonald; Andreas Suhrbier; Vasilli M Kasimov; Daniel Watterson; Nina M Pollak; David J McMillan

doi:10.1371/journal.pntd.0012352

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Development and proof of concept evaluation for a low resource compatible Chikungunya virus diagnostic

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Development and proof of concept evaluation for a low resource compatible Chikungunya virus diagnostic

Rickyle Balea, Alberto A Amarilla, Jody Hobson-Peters, Joanne Macdonald, Andreas Suhrbier, Vasilli M Kasimov, Daniel Watterson, Nina M Pollak and David J McMillan

PLoS Neglected Tropical Diseases, Vol.19(9), pp.1-15

2025

DOI: https://doi.org/10.1371/journal.pntd.0012352

PMID: 40938967

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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 Engineering
Alberto A Amarilla - The University of Queensland
Jody Hobson-Peters - The University of Queensland
Joanne Macdonald - University of the Sunshine Coast, Queensland, School of Science, Technology and Engineering
Andreas Suhrbier - QIMR Berghofer Medical Research Institute
Vasilli M Kasimov - Griffith University
Daniel Watterson - The University of Queensland
Nina M Pollak - University of the Sunshine Coast, Queensland, School of Science, Technology and Engineering
David 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
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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