Poster Presentation 12th Australasian Virology Society Meeting 2024

Integrating a chimeric Binjari virus nanotechnology into paper-based assays for POC detection of flaviviral infections in veterinary applications (#177)

Ryan Johnston 1 , Gervais Habarugira 1 2 , Sally Isberg 3 , Jessica J Harrison 1 , Mahali Morgan 1 , Lorna Melville 4 , Steven Davis 4 , Christopher Howard 5 6 , Charles Henry 7 , Paul Hick 8 , Peter Kirkland 8 , Helle Bielefeldt-Ohmann 1 5 , Roy A Hall 1 5 , Jody Hobson-Peters 1 5
  1. School of Chemistry and Molecular Biosciences, The University of Queensland, St. Lucia, Qld, Australia
  2. School of Veterinary Science, The University of Queensland, St. Lucia, Qld, Australia
  3. Centre for Crocodile Research, Noonamah, NT, Australia
  4. Department of Primary Industry and Fisheries, Katherine, NT, Australia
  5. Australian Infectious Diseases Research Centre, The University of Queensland, St. Lucia, Qld, Australia
  6. Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St. Lucia, Qld, Australia
  7. Chemical and Biological Engineering, Colorado State University, Fort Collins, Colorado, USA
  8. Elizabeth Macarthur Agricultural Institute, Menangle, NSW, Australia

Over the past five decades, flaviviruses like dengue, Zika, Japanese encephalitis virus (JEV), and West Nile virus (WNV) have increasingly spread, posing a significant, global threat to over four billion people and key livestock, including crocodiles and pigs, which are vital to Australia’s agricultural industry. The 2022 Australian JEV outbreak highlighted the urgent need for effective diagnostic tools suited to Australian conditions. Traditional lab-based diagnostics, though accurate, can be slow, cumbersome, and reliant on specialised equipment, making them less practical, particularly in remote or rural areas. Lateral flow assays (LFAs) offer a faster, simpler, and cost-effective alternative, ideal for on-site diagnostics that can be quickly deployed during outbreaks.
In this study, we enhanced LFAs for flavivirus antibody detection using antigens produced through a chimeric mosquito-specific flavivirus platform based on Binjari virus (BinJV). These BinJV-derived chimeric antigens closely mimic the epitopes on virion surfaces of native pathogenic viruses, improving antigenic accuracy. They are also safely manufactured in mosquito cell cultures, offering substantial advantages over traditional recombinant protein systems in terms of production efficiency and biosafety.
In collaboration with a consortium of industry partners, we successfully integrated BinJV/WNV and BinJV/JEV chimeras into pen-side LFAs for detecting WNV in crocodiles and JEV in pigs. In naturally exposed, farmed animals, the BinJV/WNV LFA showed 98.8% sensitivity and 100% specificity (n = 60 in farmed crocodiles), while the BinJV/JEV LFA achieved 94.8% sensitivity and 91.3% specificity (n = 93 in farmed pigs), confirming our approach's reliability. This platform provides a powerful screening tool for low-resource or remote settings, such as those in Australia. Our findings highlight the potential of BinJV-based antigens in developing LFAs to combat vector-borne diseases across multiple species, including humans, with future research expanding this technology to other flaviviruses and enhancing its industry application and field-use capabilities.