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Development and application of a tri-allelic PCR assay for screening Vgsc-L1014F kdr mutations associated with pyrethroid and organochlorine resistance in the mosquito Culex quinquefasciatus.

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SilvaMartins, Walter Fabricio, Silva Pereira, Bárbara Natieli, Vieira Alves, Ana Thayse, Murphy, Annabel, Silva Martins, Paulo Geovani, Weetman, David ORCID: https://orcid.org/0000-0002-5820-1388, Wilding, Craig Stephen and Donnelly, Martin ORCID: https://orcid.org/0000-0001-5218-1497 (2019) 'Development and application of a tri-allelic PCR assay for screening Vgsc-L1014F kdr mutations associated with pyrethroid and organochlorine resistance in the mosquito Culex quinquefasciatus.'. Parasites & Vectors, Vol 12, Issue 1, p. 232.

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Abstract

BACKGROUND
Culex quinquefasciatus has a widespread distribution across tropical and sub-tropical regions, and plays an important role in the transmission of vector-borne diseases of public health importance, including lymphatic filariasis (LF) and multiple arboviruses. Increased resistance to insecticides threatens the efficacy and sustainability of insecticide-based anti-vector interventions which mitigate the burden of mosquito transmitted diseases in endemic regions. In C. quinquefasciatus two non-synonymous voltage gated sodium channel (Vgsc) variants, both resulting in a leucine to phenylalanine change at codon 1014, are associated with resistance to pyrethroids and DDT. This tri-allelic variation has compromised the ability to perform high-throughput single-assay screening. To facilitate the detection and monitoring of the Vgsc-1014 locus in field-caught mosquitoes, an Engineered-Tail Allele-Specific-PCR (ETAS-PCR) diagnostic assay was developed and applied to wild mosquitoes from Brazil, Tanzania and Uganda.
RESULTS
This new cost-effective, single-tube assay was compared to two, well-established, genotyping approaches, pyrosequencing and TaqMan. The ETAS-PCR assay showed high specificity for discriminating the three alleles at Vgsc-L1014F, with genotyping results strongly correlated with pyrosequencing and TaqMan results (98.64% and 100% respectively).
CONCLUSIONS
Our results support the utility of the ETAS-PCR/Vgsc-1014 diagnostic assay, which stands as an effective alternative for genotyping tri-allelic variants.

Item Type: Article
Subjects: QU Biochemistry > Genetics > QU 475 Genetic processes
QU Biochemistry > Genetics > QU 550 Genetic techniques. PCR. Chromosome mapping
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
WA Public Health > Preventive Medicine > WA 240 Disinfection. Disinfestation. Pesticides (including diseases caused by)
Faculty: Department: Biological Sciences > Vector Biology Department
Digital Object Identifer (DOI): https://doi.org/10.1186/s13071-019-3490-z
Depositing User: Stacy Murtagh
Date Deposited: 16 May 2019 13:22
Last Modified: 16 Sep 2019 09:18
URI: https://archive.lstmed.ac.uk/id/eprint/10784

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