RT-qPCR mosquito arbovirus screening results, 2017-2021, South Nation River watershed

<p> Pool-by-pool RT-qPCR mosquito arbovirus screening results (.xlsx), 2017-2021, South Nation River watershed. Viruses tested include: California serogroup viruses (CSGv), West Nile virus (WNv), eastern equine encephelitis virus (EEEv), and Cache Valley virus (CVv).<b> Includes a sheet with column descriptions.</b></p> <br>

<h3> Methods </h3> <br>

<p> Between 2017 and 2021 (excluding 2020 due to COVID-19), mosquitoes were collected across semi-urban and rural habitats of the South Nation River watershed, eastern Ontario. Pools of 1–50 females per species (≤3 pools, max. 150 individuals) were tested, with some Ceratopogonidae also included. Samples from 2017–2019 were homogenized at the National Microbiology Laboratory (NML) as described in Avramov et al. (1) and RNA-extracted with the QIAamp Viral Mini Kit (QIAGEN™) with minor modifications (2). In 2021, homogenization was performed at Carleton University under the same conditions (1), with RNA extraction by high-throughput phenol–chloroform (1). RT-qPCR screening for CSGv used Wang et al.’s primers/probes (3,4) at NML (2017–2018) and Carleton (2019, 2021). WNv and EEEv were screened at NML (2017–2019) and Carleton (2021) with validated primers/probes (5,6). CVv was screened only at NML in 2017–2018 using Wang et al. (3). Virus-positive pools processed at Carleton underwent RNA re-extraction with the QIAamp Viral Mini Kit, followed by replicate RT-qPCR; Cq < 38 in both replicates defined a positive. All procedures followed institutional biosafety standards.</p>

<h3>References</h3>
<ol>
<li>Avramov M, Gallo V, Gross A, Lapen DR, Ludwig A, Cullingham CI. A cost-effective RNA extraction and RT-qPCR approach to detect California serogroup viruses from pooled mosquito samples. Sci Rep. 2024 Jan 29;14(1):2339. </li>
<li>Villeneuve CA, Buhler KJ, Iranpour M, Avard E, Dibernardo A, Fenton H, et al. New records of California serogroup viruses in Aedes mosquitoes and first detection in simulioidae flies from Northern Canada and Alaska. Polar Biol. 2021 Sep 1;44(9):1911–5.</li>
<li>Wang H, Nattanmai S, Kramer LD, Bernard KA, Tavakoli NP. A duplex real-time reverse transcriptase polymerase chain reaction assay for the detection of California serogroup and Cache Valley viruses. Diagn Microbiol Infect Dis. 2009 Oct 1;65(2):150–7.</li>
<li> Buhler KJ, Dibernardo A, Pilfold NW, Harms NJ, Fenton H, Carriere S, et al. Widespread Exposure to Mosquitoborne California Serogroup Viruses in Caribou, Arctic Fox, Red Fox, and Polar Bears, Canada. Emerg Infect Dis. 2023 Jan;29(1):54–63.</li>
<li> Lanciotti RS, Kerst AJ, Nasci RS, Godsey MS, Mitchell CJ, Savage HM, et al. Rapid Detection of West Nile Virus from Human Clinical Specimens, Field-Collected Mosquitoes, and Avian Samples by a TaqMan Reverse Transcriptase-PCR Assay. J Clin Microbiol. 2000 Nov;38(11):4066–71.</li>
<li> Lambert AJ, Martin DA, Lanciotti RS. Detection of North American Eastern and Western Equine Encephalitis Viruses by Nucleic Acid Amplification Assays. J Clin Microbiol. 2003 Jan;41(1):379–85.</li>
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