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Detection and quantitation of copy number variation in the voltage-gated sodium channel gene of the mosquito Culex quinquefasciatus

Martins, WFS, Subramaniam, K, Steen, K, Mawejje, H, Liloglou, T, Donnelly, MJ and Wilding, CS (2017) Detection and quantitation of copy number variation in the voltage-gated sodium channel gene of the mosquito Culex quinquefasciatus. Scientific Reports, 7 (1). pp. 1-10. ISSN 2045-2322

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Insecticide resistance is typically associated with alterations to the insecticidal target-site or with gene expression variation at loci involved in insecticide detoxification. In some species copy number variation (CNV) of target site loci (e.g. the Ace-1 target site of carbamate insecticides) or detoxification genes has been implicated in the resistance phenotype. We show that field-collected Ugandan
Culex quinquefasciatus display CNV for the voltage-gated sodium channel gene (Vgsc), target-site of pyrethroid and organochlorine insecticides. In order to develop field-applicable diagnostics for Vgsc
CN, and as a prelude to investigating the possible association of CN with insecticide resistance, three
assays were compared for their accuracy in CN estimation in this species. The gold standard method is
droplet digital PCR (ddPCR), however, the hardware is prohibitively expensive for widespread utility.
Here, ddPCR was compared to quantitative PCR (qPCR) and pyrosequencing. Across all platforms, CNV was detected in ≈10% of mosquitoes, corresponding to three or four copies (per diploid genome).
ddPCR and qPCR-Std-curve yielded similar predictions for Vgsc CN, indicating that the qPCR protocol developed here can be applied as a diagnostic assay, facilitating monitoring of Vgsc CN in wild populations and the elucidation of association between the Vgsc CN and insecticide resistance.

Item Type: Article
Subjects: Q Science > QH Natural history > QH301 Biology
Q Science > QH Natural history > QH426 Genetics
Divisions: Natural Sciences & Psychology (closed 31 Aug 19)
Publisher: Nature Publishing Group
Date Deposited: 20 Jul 2017 14:19
Last Modified: 04 Sep 2021 11:20
DOI or ID number: 10.1038/s41598-017-06080-8
URI: https://researchonline.ljmu.ac.uk/id/eprint/6836
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