Genomic and Molecular Dataset on Target-Site Mutation Genotyping of Insecticide-Resistant Anopheles gambiae Populations in Ota
Description
This dataset provides a comprehensive molecular and genomic characterisation of insecticide-resistant Anopheles gambiae populations collected across five breeding sites in Ota, Ogun State, Nigeria.
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The dataset was generated through a structured entomological and molecular workflow designed to characterise target-site resistance mutations in Anopheles gambiae populations from Ota, Ogun State, Nigeria. Mosquito larvae were collected between January and July 2025 from five breeding sites—Nestle, Chelsea, Canaan City, Atan and Iju—using standard dipping techniques. Collected larvae were transported to the Covenant University Insectary and reared under controlled temperature, humidity and light conditions. Adults were morphologically identified using standard African Anopheles taxonomic keys, after which molecular species confirmation was performed using the ribosomal DNA intergenic spacer PCR method. Genomic DNA was extracted from individual adult females using the Livak non-organic extraction protocol, which yields DNA suitable for PCR-based genotyping and sequencing. Insecticide resistance was assessed using WHO susceptibility tube assays with 0.75% permethrin and 0.1% bendiocarb papers sourced from the WHO Collaborating Centre. Batches of 20–25 non-blood-fed females aged 3–5 days were exposed for 60 minutes, and mortality was recorded after 24 hours following the WHO 2016 guidelines.Target-site mutations (L1014F and L1014S) in the voltage-gated sodium channel gene were detected using allele-specific PCR with validated primers. PCR amplification was performed using OneTaq DNA Polymerase under optimised cycling conditions, and the products were resolved on 1.5% agarose gels stained with SYBR Safe and visualised on a Bio-Rad Gel Doc system. Representative amplicons were purified using QIAquick kits and sequenced via Sanger sequencing at Inqaba Biotechnical Industries. Sequence chromatograms were manually inspected, trimmed, and aligned using ClustalW, implemented in MEGA version 12. Phylogenetic analyses were conducted using the Maximum Likelihood method with 1,000 bootstraps. Genetic diversity indices, including haplotype diversity and nucleotide diversity, were calculated using DnaSP version 6. Population differentiation (F_ST) and gene flow (Nm) were estimated using R studio. Principal Component Analysis for population clustering was performed using R studio.
Institutions
- Covenant University