A next generation targeted amplicon sequencing method to screen for insecticide resistance mutations in Aedes aegypti populations reveals a rdl mutation in mosquitoes from Cabo Verde.

Publisher: Public Library of Science Country of Publication: United States NLM ID: 101291488 Publication Model: eCollection Cited Medium: Internet ISSN: 1935-2735 (Electronic) Linking ISSN: 19352727 NLM ISO Abbreviation: PLoS Negl Trop Dis Subsets: MEDLINE

Original Publication: San Francisco, CA : Public Library of Science

Aedes*/genetics ; Insecticides*/pharmacology ; Pyrethrins*/pharmacology ; Voltage-Gated Sodium Channels*/genetics ; Zika Virus* ; Zika Virus Infection*; Animals ; Insecticide Resistance/genetics ; DDT ; Cabo Verde ; Mosquito Vectors/genetics ; Mutation

Aedes mosquito vectors transmit many viruses of global health concern, including dengue, chikungunya and Zika. These vector-borne viral diseases have a limited number of treatment options, and vaccines vary in their effectiveness. Consequently, integrated vector management is a primary strategy for disease control. However, the increasing emergence and spread of insecticide resistance is threatening the efficacy of vector control methods. Identifying mutations associated with resistance in vector populations is important to monitor the occurrence and evolution of insecticide resistance and inform control strategies. Rapid and cost-effective genome sequencing approaches are urgently needed. Here we present an adaptable targeted amplicon approach for cost-effective implementation within next generation sequencing platforms. This approach can identify single nucleotide polymorphisms (SNPs) and small insertions and deletions (indels) in genes involved in insecticide resistance in Aedes aegypti mosquitoes. We designed and tested eleven amplicons, which included segments of the ace-1 (carbamate target), the Voltage-Gated Sodium Channel (vgsc; pyrethroids, DDT and organochlorines), and rdl (dieldrin) genes; thereby covering established knockdown resistance (kdr) mutations (e.g., S989P, I1011M/V, V1016G/I and F1534C), with the potential to identify novel ones. The amplicon assays were designed with internal barcodes, to facilitate multiplexing of large numbers of mosquitoes at low cost, and were sequenced using an Illumina platform. Our approach was evaluated on 152 Ae. aegypti mosquitoes collected in Cabo Verde, an archipelago with a history of arbovirus outbreaks. The amplicon sequence data revealed 146 SNPs, including four non-synonymous polymorphisms in the vgsc gene, one in ace-1 and the 296S rdl mutation previously associated with resistance to organochlorines. The 296S rdl mutation was identified in 98% of mosquitoes screened, consistent with the past use of an organochlorine compound (e.g., DDT). Overall, our work shows that targeted amplicon sequencing is a rapid, robust, and cost-effective tool that can be used to perform high throughput monitoring of insecticide resistance.
Competing Interests: The authors have declared that no competing interests exist.
(Copyright: © 2022 Collins et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

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MR/M01360X/1 United Kingdom MRC_ Medical Research Council; MR/N010469/1 United Kingdom MRC_ Medical Research Council; MR/R025576/1 United Kingdom MRC_ Medical Research Council; MR/R020973/1 United Kingdom MRC_ Medical Research Council; MR/M01360X/1 United Kingdom MRC_ Medical Research Council; MR/R025576/1 United Kingdom MRC_ Medical Research Council; MR/R020973/1 United Kingdom MRC_ Medical Research Council; United Kingdom BB_ Biotechnology and Biological Sciences Research Council

CIW5S16655 (DDT)
0 (Insecticides)
0 (Pyrethrins)
0 (Voltage-Gated Sodium Channels)

Date Created: 20221213 Date Completed: 20221215 Latest Revision: 20221224

20231215

PMC9746995

10.1371/journal.pntd.0010935

36512510