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Martins, Fabricio, Subramaniam, Krishanthi, Steen, Keith 
ORCID: https://orcid.org/0000-0002-8933-8643, Mawejje, Henry, Liloglou, Triantafillos, Donnelly, Martin ORCID: https://orcid.org/0000-0001-5218-1497 and Wilding, Craig Stephen
(2017)
'Detection and quantitation of copy number variation in the voltage-gated sodium channel gene of the mosquito Culex quinquefasciatus'. Scientific Reports, Vol 19, Issue 7 (1), p. 5821.
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Scientific_Reports_Martins et al 2017 CNV in Culex.pdf - Published Version Available under License Creative Commons Attribution. Download (1MB) | Preview |
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 |
|---|---|
| Subjects: | QU Biochemistry > Genetics > QU 450 General Works QU Biochemistry > Genetics > QU 460 Genomics. Proteomics QX Parasitology > Insects. Other Parasites > QX 510 Mosquitoes QX Parasitology > Insects. Other Parasites > QX 530 Culex QX Parasitology > Insects. Other Parasites > QX 600 Insect control. Tick control QX Parasitology > Insects. Other Parasites > QX 650 Insect vectors |
| Faculty: Department: | Biological Sciences > Vector Biology Department |
| Digital Object Identifer (DOI): | https://doi.org/10.1038/s41598-017-06080-8 |
| Depositing User: | Stacy Murtagh |
| Date Deposited: | 01 Aug 2017 15:57 |
| Last Modified: | 16 Sep 2019 09:17 |
| URI: | https://archive.lstmed.ac.uk/id/eprint/7432 |
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