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Rapid molecular assays for the detection of the four dengue viruses in infected mosquitoes [version 1; peer review: 1 approved]
Preprint   Open access

Rapid molecular assays for the detection of the four dengue viruses in infected mosquitoes [version 1; peer review: 1 approved]

Madeeha Ahmed, Nina Pollak, Leon E Hugo, Andrew F van den Hurk, Jody Hobson-Peters and Joanne Macdonald
Gates Open Research, Vol.6, pp.1-13
F1000 Research Ltd
2022
PMCID: PMC9816563
PMID: 36636741
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Rapid molecular assays for the detection of the four dengue viruses in infected mosquitoes version 1 peer review - 1 approved1.84 MBDownloadView
Preprint Version Open Access CC BY V4.0
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https://doi.org/10.12688/gatesopenres.13534.1View
Preprint Version

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Abstract

dengue virus recombinase olymerase amplification isothermal amplification lateral flow detection rapid molecular assays mosquitoes Aedes aegypti mosquito surveillance
The pantropic emergence of severe dengue disease can partly be attributed to the co-circulation of different dengue viruses (DENVs) in the same geographical location. Effective monitoring for circulation of each of the four DENVs is critical to inform disease mitigation strategies. In low resource settings, this can be effectively achieved by utilizing inexpensive, rapid, sensitive and specific assays to detect viruses in mosquito populations. In this study, we developed four rapid DENV tests with direct applicability for low-resource virus surveillance in mosquitoes. The test protocols utilize a novel sample preparation step, a single-temperature isothermal amplification, and a simple lateral flow detection. Analytical sensitivity testing demonstrated tests could detect down to 1,000 copies/µL of virus-specific DENV RNA, and analytical specificity testing indicated tests were highly specific for their respective virus, and did not detect closely related flaviviruses. All four DENV tests showed excellent diagnostic specificity and sensitivity when used for detection of both individually infected mosquitoes and infected mosquitoes in pools of uninfected mosquitoes. With individually infected mosquitoes, the rapid DENV-1,-2 and-3 tests showed 100% diagnostic sensitivity (95% CI = 69% to 100%, n=8 for DENV-1; n=10 for DENV 2,3) and the DENV-4 test showed 92% diagnostic sensitivity (CI: 62% to 100%, n=12) along with 100% diagnostic specificity (CI: 48–100%) for all four tests. Testing infected mosquito pools, the rapid DENV-2,-3 and-4 tests showed 100% diagnostic sensitivity (95% CI = 69% to 100%, n=10) and the Open Peer Review Approval Status AWAITING PEER REVIEW

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