West Nile virus in crocodiles and mosquitoes in Zimbabwe

Roy, Mohashweta; Mufunda, F.; Slawski, D.; Mutswiri, C.; Mvere, I.; Slawski, K.; Bezuidenhout, Abraham J.; Piras, Ilaria M.; Hanyire, T.; Markey, Bryan; Barry, Gerald

doi:10.1101/2021.02.19.431612

Cited by 2 publications

(1 citation statement)

References 9 publications

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“…Similar conditions in previous years have also resulted in outbreaks of WNV in horses (Frost et al 2012; Read et al 2019). Another key industry affected by WNV, not only in Australia, but elsewhere in the world is the commercial crocodilian farming sector (Isberg et al 2019; Jacobson et al 2005; Roy et al 2021). The Australian endemic strain of WNV, Kunjin (WNV KUN ) currently causes substantial financial losses in the commercial crocodile-farming sector due to hide imperfections as a result of infections (Habarugira et al 2020a; Isberg et al 2019).…”

Section: Introductionmentioning

confidence: 99%

Application of chimeric antigens to paper-based diagnostics for detection of West Nile virus infections ofCrocodylus porosus –a novel animal test case

Johnston,

Habarugira,

Harrison

et al. 2024

Preprint

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Laboratory-based diagnostics like plaque reduction neutralization tests (PRNT) and ELISA are commonly used to detect seroconversion to flavivirus infections. However, faster, qualitative screening methods are needed for quicker diagnosis and better patient outcomes. Lateral flow assays (LFAs) can provide rapid results (5-15 mins) at the point-of-care, yet few commercial flavivirus antibody detection LFAs are available. We developed an LFA using novel chimeric viral antigens produced by genetically modifying the mosquito restricted Binjari virus (BinJV) to display the outer virion proteins of pathogenic viruses such as West Nile virus (WNV). The BinJV chimeric platform offers various advantages for diagnostic assay development, including rapid construction of new chimeras in response to emerging viral variants, safe, scalable antigen manufacturing, and structural indistinguishability to the wild-type pathogenic virion. As a demonstration of feasibility, we applied chimeric WNV (BinJV/WNV) antigen to LFA as the capture/test line reagent for detection of seroconversion of crocodilians to WNV – a virus affecting crocodilians on multiple continents. We verified the antigenic conservation of the chimera when applied to the LFA detection surface using monoclonal antibodies. Using well-characterised sera (n=60) from WNV seropositive or flavivirus naive Australian saltwater crocodiles (Crocodylus porosus), we illustrated 100% sensitivity and specificity, with results achieved in less than 15 minutes. The LFA further accurately detected seroconversion in animals experimentally infected with WNV. This qualitative screening method can be performed both inside and outside of a laboratory, and the assay design will guide the optimization of similar tests for vector borne virus infection detection in both humans and other animals.

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