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Silva Martins, WF, Silva Pereira, BN, Vieira Alves, AT, Murphy, A, Silva Martins, PG, Weetman, D, Wilding, CS and Donnelly, MJ (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 and Vectors, 12 (1). ISSN 1756-3305
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Martins et al 2019 Triallelic kdr PCR in Culex.pdf - Published Version Available under License Creative Commons Attribution. Download (962kB) | Preview |
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 |
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| Uncontrolled Keywords: | 1108 Medical Microbiology, 1117 Public Health and Health Services |
| Subjects: | Q Science > QR Microbiology R Medicine > RA Public aspects of medicine > RA0421 Public health. Hygiene. Preventive Medicine R Medicine > RM Therapeutics. Pharmacology |
| Divisions: | Natural Sciences & Psychology (closed 31 Aug 19) |
| Publisher: | BioMed Central |
| Date Deposited: | 20 May 2019 09:51 |
| Last Modified: | 04 Sep 2021 09:23 |
| DOI or ID number: | 10.1186/s13071-019-3490-z |
| URI: | https://researchonline.ljmu.ac.uk/id/eprint/10727 |
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