Detection and quantitation of copy number variation in the voltagegated sodium channel gene of the mosquito <i>Culex quinquefasciatus</i>

Martins, Walter Fabricio Silva, Subramaniam, Krishanthi ORCID: 0000-0002-1734-9351, Steen, Keith, Mawejje, Henry, Liloglou, Triantafillos ORCID: 0000-0003-0460-1404, Donnelly, Martin James and Wilding, Craig Stephen (2017) Detection and quantitation of copy number variation in the voltagegated sodium channel gene of the mosquito <i>Culex quinquefasciatus</i>. SCIENTIFIC REPORTS, 7 (1). 5821-.

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Official URL: http://dx.doi.org/10.1038/s41598-017-06080-8

Abstract

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
Uncontrolled Keywords:	Animals, Culex, Reproducibility of Results, Gene Dosage, Haplotypes, Genes, Insect, DNA Copy Number Variations, Voltage-Gated Sodium Channels
Depositing User:	Symplectic Admin
Date Deposited:	27 Jul 2017 10:41
Last Modified:	12 Oct 2023 16:57
DOI:	10.1038/s41598-017-06080-8
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3008662