P-3 - Adaptation and Optimization of a Triplex Reverse Transcription Polymerase Chain Reaction Protocol for Arbovirus Surveillance in Canyon County Idaho

Abstract: In 2023 the Canyon County Mosquito Abatement District upgraded their arbovirus surveillance program from relying on RAMP testing for West Nile Virus (WNV) and testing through the Idaho Bureau of Laboratories (IBL) for Western Equine Encephalitis Virus (WEEV) and St. Louis Encephalitis Virus (SLEV) in mosquito pools to a triplex RT-qPCR utilizing Co-Diagnostics Vector Smart North American Mosquito West (NAM-w) multiplex assay for the detection of WNV, WEEV, and SLEV. Initial adaptation of the NAM-w assay in 2023 resulted in concerns for the validity of our mosquito pool virus detection results obtained during the 2023 season when tested in duplicate through IBL. To address those concerns in 2024 an alternate protocol was adapted, optimized for our equipment, and implemented in our arbovirus surveillance program. Results from our adaptation of this protocol during the 2024 mosquito season will be presented.