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April 10, 2026Pathogens0 citationsOpen Access

From RAMP to Triplex RT-qPCR: Modernizing Arbovirus Surveillance and Confirming the First Aedes aegypti in Idaho

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HWHeather M. WardJLJames J. LundersCOChris Ocegueda

Key Points

  • The aim is to modernize arbovirus surveillance and enhance mosquito identification capacities in Idaho.
  • Implemented triplex RT-qPCR for detecting WNV, WEEV, and SLEV.
  • Consolidated and geospatially harmonized historical trapping datasets for multi-year comparisons.
  • Standardized mosquito identification and pooling procedures.
  • Conducted routine gravid trap surveillance for Aedes aegypti.
  • Detectable WNV activity was observed each year, peaking in 2023 with 192 positive pools.
  • No detections of WEEV or SLEV during the study period.
  • Enhanced laboratory capacity allowed for same-day reporting and decreased processing times.

Abstract

West Nile virus (WNV) remains the most frequently reported locally acquired arboviral infection in the United States, yet many small and mid-sized mosquito abatement districts lack the diagnostic capacity and integrated data systems needed for rapid detection and response. The Canyon County Mosquito Abatement District (CCMAD) in southwestern Idaho undertook a multi-year capacity-building effort to expand arbovirus surveillance, standardize mosquito identification and pooling procedures, and implement triplex RT-qPCR testing for WNV, Western equine encephalitis virus (WEEV), and St. Louis encephalitis virus (SLEV). Historical trapping datasets (2021–2025) were consolidated, geospatially harmonized, and grouped into biologically meaningful sampling units to enable multi-year spatial comparisons. Surveillance revealed recurrent WNV activity annually, with peak transmission occurring between epidemiological weeks 31 and 37. The highest WNV activity occurred in 2023 and 2025, with 192 and 92 positive pools, respectively, while no WEEV or SLEV detections were observed. Enhanced laboratory capacity reduced sample-processing times, decreased the reliance on external confirmatory testing, lowered per-pool testing costs, and enabled same-day reporting to operational staff. In 2025, routine gravid trap surveillance detected a single Aedes aegypti, which was identified morphologically and subsequently confirmed by DNA barcoding, prompting targeted follow-up trapping. CCMAD’s integrated approach provides a scalable model for strengthening local surveillance and response capabilities in resource-limited settings.

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Cite This Study

Ward et al. (2026) studied this question.

synapsesocial.com/papers/69d8967d6c1944d70ce07ef4https://doi.org/10.3390/pathogens15040406
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