Rapid determination of infection in mosquitoes is currently performed using real-time polymerase chain reaction, a laboratory-based procedure that requires thermocycling equipment and takes several hours to complete. We identified recombinase polymerase amplification (RPA) coupled with lateral flow detection as a simple protocol amenable to field-based detection, as the assay is supplied in freeze-dried ampoules and requires only heating at 39 °C for operation. We investigated use of this technology for detection of viral RNA from Japanese encephalitis, Murray Valley encephalitis, and West Nile viruses. The assays can differentiate each virus from other closely related pathogens, and can be performed in less than 30 minutes. We also developed a novel multiplex lateral flow strip that provides a simple text display output for potentially simplified diagnosis. The output text display requires no batteries or wires because the molecules themselves power the device, and reduces the cost of the assay compared to performing multiple individual reactions. This technology workflow provides a potentially simplified and efficient protocol for detection, and can readily be applied for detection of other arboviruses.
12th Symposium of the Mosquito Control Association of Australia and Arbovirus Research in Australia: Managing challenges and threats with new technology and research, Surfers Paradise, Australia 4-9 September 2016
12th Symposium of the Mosquito Control Association of Australia and Arbovirus Research in Australia Book of Abstracts / pp.8