{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,8]],"date-time":"2026-04-08T13:00:14Z","timestamp":1775653214090,"version":"3.50.1"},"reference-count":44,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2026,4,8]],"date-time":"2026-04-08T00:00:00Z","timestamp":1775606400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Idaho Department of Health and Welfare Division of Public Health Mosquito Abatement District contracts"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>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\u20132025) 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\u2019s integrated approach provides a scalable model for strengthening local surveillance and response capabilities in resource-limited settings.<\/jats:p>","DOI":"10.3390\/pathogens15040406","type":"journal-article","created":{"date-parts":[[2026,4,8]],"date-time":"2026-04-08T11:41:18Z","timestamp":1775648478000},"page":"406","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["From RAMP to Triplex RT-qPCR: Modernizing Arbovirus Surveillance and Confirming the First Aedes aegypti in Idaho"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3939-5308","authenticated-orcid":false,"given":"Heather M.","family":"Ward","sequence":"first","affiliation":[{"name":"Canyon County Mosquito Abatement District, Nampa, ID 83686, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"James J.","family":"Lunders","sequence":"additional","affiliation":[{"name":"Canyon County Mosquito Abatement District, Nampa, ID 83686, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chris","family":"Ocegueda","sequence":"additional","affiliation":[{"name":"Canyon County Mosquito Abatement District, Nampa, ID 83686, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"308","DOI":"10.1001\/jama.2013.8042","article-title":"West Nile Virus: Review of the Literature","volume":"310","author":"Petersen","year":"2013","journal-title":"JAMA"},{"key":"ref_2","unstructured":"Centers for Disease Control and Prevention (CDC) (2003). West Nile virus activity\u2014United States, November 20\u201325, 2003. MMWR Morb. Mortal. Wkly. Rep., 52, 1060."},{"key":"ref_3","unstructured":"Idaho Department of Health and Welfare (IDHW) (2003). Idaho Reports Its First Human Case of West Nile Fever, Idaho Disease Bulletin."},{"key":"ref_4","unstructured":"Idaho Department of Health and Welfare (IDHW) (2023). West Nile Virus Concatenated File 2003-2023."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"423","DOI":"10.7589\/0090-3558-16.3.423","article-title":"A serologic survey for selected infectious diseases of black bears in idaho","volume":"16","author":"Binninger","year":"1980","journal-title":"J. Wildl. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Swetnam, D.M., Stuart, J.B., Young, K., Maharaj, P.D., Fang, Y., Garcia, S., Barker, C.M., Smith, K., Godsey, M.S., and Savage, H.M. (2020). Movement of St. Louis Encephalitis Virus in the Western United States, 2014\u20132018. PLoS Negl. Trop. Dis., 14.","DOI":"10.1371\/journal.pntd.0008343"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2079","DOI":"10.3390\/v5092079","article-title":"Ecology of West Nile Virus in North America","volume":"5","author":"Reisen","year":"2013","journal-title":"Viruses"},{"key":"ref_8","unstructured":"CDC (2023). HHS West Nile Virus Surveillance and Control Guidelines."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Snyder, R.E., Feiszli, T., Foss, L., Messenger, S., Fang, Y., Barker, C.M., Reisen, W.K., Vugia, D.J., Padgett, K.A., and Kramer, V.L. (2020). West Nile Virus in California, 2003\u20132018: A Persistent Threat. PLoS Negl. Trop. Dis., 14.","DOI":"10.1371\/journal.pntd.0008841"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"938","DOI":"10.15585\/mmwr.mm6928a6","article-title":"Continuation of Mosquito Surveillance and Control During Public Health Emergencies and Natural Disasters","volume":"69","author":"Connelly","year":"2020","journal-title":"MMWR Morb. Mortal. Wkly. Rep."},{"key":"ref_11","unstructured":"National Association of County and City Health Officials (NACCHO) (2017). Mosquito Control Capabilities in the United States, NACCHO."},{"key":"ref_12","unstructured":"National Association of County and City Health Officials (NACCHO) (2020). Vector Control at the Local Level: Findings from the 2020 Vector Control Assessment, NACCHO."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"21","DOI":"10.2987\/13-6386.1","article-title":"Evaluation of a Rapid Analyte Measurement Platform and Real-Time Reverse-Transcriptase Polymerase Chain Reaction Assay West Nile Virus Detection System in Mosquito Pools","volume":"30","author":"Burkhalter","year":"2014","journal-title":"J. Am. Mosq. Control Assoc."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1420","DOI":"10.1093\/jme\/tjx088","article-title":"Updated Reported Distribution of Aedes (Stegomyia) aegypti and Aedes (Stegomyia) albopictus (Diptera: Culicidae) in the United States, 1995\u20132016","volume":"54","author":"Hahn","year":"2017","journal-title":"J. Med. Entomol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e08347","DOI":"10.7554\/eLife.08347","article-title":"The Global Distribution of the Arbovirus Vectors Aedes Aegypti and Ae. Albopictus","volume":"4","author":"Kraemer","year":"2015","journal-title":"eLife"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Burkett-Cadena, N.D. (2014). Mosquitoes of the Southeastern United States, University of Alabama Press.","DOI":"10.2307\/jj.30347333"},{"key":"ref_17","unstructured":"Carpenter, S.J., and LaCasse, W.J. (1974). Mosquitoes of North America (North of Mexico), University of California Press."},{"key":"ref_18","unstructured":"Darsie, R.F., and Ward, R.A. (2016). Identification and Geographical Distribution of the Mosquitoes of North America, North of Mexico, University Press of Florida."},{"key":"ref_19","unstructured":"Harrison, B.A., Byrd, B.D., Sither, C.B., and Whitt, P.B. (2016). The Mosquitoes of the Mid-Atlantic Region: An Identification Guide, Western Carolina University."},{"key":"ref_20","unstructured":"Rose, D.A., Kondratieff, B.C., and Weissmann, M.J. (2017). Colorado Mosquitoes (Diptera: Culicidae). Insects of Western North America; Contributions of the C.P. Gillette Museum of Arthropod Diversity, Department of Bioagricultural Sciences and Pest Management, Colorado State University."},{"key":"ref_21","first-page":"440","article-title":"Identification of Damaged Adult Female Specimens of Aedes Albopictus and Aedes Aegypti in the New World","volume":"10","author":"Savage","year":"1994","journal-title":"J. Am. Mosq. Control Assoc."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"359","DOI":"10.4269\/ajtmh.2012.11-0662","article-title":"Evaluation of a Rapid Analyte Measurement Platform for West Nile Virus Detection Based on United States Mosquito Control Programs","volume":"87","author":"Kesavaraju","year":"2012","journal-title":"Am. Soc. Trop. Med. Hyg."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1093\/jme\/tjv021","article-title":"Multiplex qRT-PCR for the Detection of Western Equine Encephalomyelitis, St. Louis Encephalitis, and West Nile Viral RNA in Mosquito Pools (Diptera: Culicidae)","volume":"52","author":"Brault","year":"2015","journal-title":"J. Med. Entomol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"4066","DOI":"10.1128\/JCM.38.11.4066-4071.2000","article-title":"Rapid Detection of West Nile Virus from Human Clinical Specimens, Field-Collected Mosquitoes, and Avian Samples by a TaqMan Reverse Transcriptase-PCR Assay","volume":"38","author":"Lanciotti","year":"2000","journal-title":"J. Clin. Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1264","DOI":"10.1128\/JCM.39.4.1264-1271.2001","article-title":"High-Throughput Detection of West Nile Virus RNA","volume":"39","author":"Shi","year":"2001","journal-title":"J. Clin. Microbiol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"657","DOI":"10.1089\/vbz.2011.0786","article-title":"National and Regional Associations between Human West Nile Virus Incidence and Demographic, Landscape, and Land Use Conditions in the Coterminous United States","volume":"12","author":"DeGroote","year":"2012","journal-title":"Vector Borne Zoonotic Dis. Larchmt. N"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1450","DOI":"10.1603\/033.046.0630","article-title":"Seasonal Variation of Potential Flavivirus Vectors in an Urban Biological Reserve in Northeastern Brazil","volume":"46","author":"Medeiros","year":"2009","journal-title":"J. Med. Entomol."},{"key":"ref_28","first-page":"40","article-title":"Ecological Factors Associated with West Nile Virus Transmission, Northeastern United States","volume":"5","author":"Brown","year":"2008","journal-title":"EcoHealth"},{"key":"ref_29","first-page":"294","article-title":"DNA Primers for Amplification of Mitochondrial Cytochrome c Oxidase Subunit I from Diverse Metazoan Invertebrates","volume":"3","author":"Folmer","year":"1994","journal-title":"Mol. Mar. Biol. Biotechnol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1186\/s13071-014-0569-4","article-title":"DNA Barcoding: Complementing Morphological Identification of Mosquito Species in Singapore","volume":"7","author":"Chan","year":"2014","journal-title":"Parasit. Vectors"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1016\/S0022-2836(05)80360-2","article-title":"Basic local alignment search tool","volume":"215","author":"Altschul","year":"1990","journal-title":"J. Mol. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"D123","DOI":"10.1093\/nar\/gkad1044","article-title":"Database resources of the National Center for Biotechnology Information","volume":"52","author":"Sayers","year":"2024","journal-title":"Nucleic Acids Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1111\/j.1471-8286.2007.01678.x","article-title":"BOLD: The Barcode of Life Data System (http:\/\/www.boldsystems.org)","volume":"7","author":"Ratnasingham","year":"2007","journal-title":"Mol. Ecol. Notes"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"437","DOI":"10.1186\/s13071-022-05494-2","article-title":"Modification of the Folmer Primers for the Cytochrome c Oxidase Gene Facilitates Identification of Mosquitoes","volume":"15","author":"Hoque","year":"2022","journal-title":"Parasit. Vectors"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Campbell, L.P. (2015). Climate Change Influences on Global Distributions of Aedes Aegypti and Aedes Albopictus. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0137851"},{"key":"ref_36","first-page":"193","article-title":"Evaluation of Automated Nucleic Acid Extraction and Multiplex Real-Time RT-PCR for Arbovirus Surveillance","volume":"168","author":"Dyer","year":"2010","journal-title":"J. Virol. Methods"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"487","DOI":"10.1128\/CMR.00072-15","article-title":"Zika Virus","volume":"29","author":"Musso","year":"2016","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1007\/s00430-023-00762-z","article-title":"Detection of DENV-2 and ZIKV Coinfection in Southeastern Brazil by Serum and Urine Testing","volume":"212","year":"2023","journal-title":"Med. Microbiol. Immunol. (Berl.)"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1111\/nyas.14957","article-title":"Positive-Strand RNA Viruses\u2014A Keystone Symposia Report","volume":"1521","author":"Cable","year":"2023","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1186\/s40249-016-0148-3","article-title":"Outwitting Dengue Threat and Epidemics Resurgence in Asia-Pacific Countries: Strengthening Integrated Dengue Surveillance, Monitoring and Response Systems","volume":"5","author":"Tambo","year":"2016","journal-title":"Infect. Dis. Poverty"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"466","DOI":"10.1016\/j.aogh.2015.02.006","article-title":"The Burden of Dengue and Chikungunya Worldwide: Implications for the Southern United States and California","volume":"80","author":"Fredericks","year":"2015","journal-title":"Ann. Glob. Health"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Mulderij-Jansen, V. (2022). Effectiveness of Aedes-Borne Infectious Disease Control in Latin America and the Caribbean Region: A Scoping Review. PLoS ONE, 17.","DOI":"10.1371\/journal.pone.0277038"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1186\/s40409-018-0162-y","article-title":"Yellow Fever in Africa and the Americas: A Historical and Epidemiological Perspective","volume":"24","author":"Chippaux","year":"2018","journal-title":"J. Venom. Anim. Toxins Trop. Dis."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"4506","DOI":"10.1128\/JCM.39.12.4506-4513.2001","article-title":"Nucleic Acid Sequence-Based Amplification Assays for Rapid Detection of West Nile and St. Louis Encephalitis Viruses","volume":"39","author":"Lanciotti","year":"2001","journal-title":"J. Clin. Microbiol."}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/15\/4\/406\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,8]],"date-time":"2026-04-08T11:55:27Z","timestamp":1775649327000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/15\/4\/406"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,8]]},"references-count":44,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2026,4]]}},"alternative-id":["pathogens15040406"],"URL":"https:\/\/doi.org\/10.3390\/pathogens15040406","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,8]]}}}