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.

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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