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ViewAttendees 4

10-Minute Paper

Medical, Urban, and Veterinary Entomology

MUVE: Vector Control and Resistance Management

A molecular inversion probe panel for high-throughput insecticide resistance monitoring of malaria vectors

Wednesday, November 8, 2023
8:12 AM – 8:24 AM ET
Location: Gaylord National Resort & Convention Center, National Harbor 2
In-Person
Insecticide resistance is expanding throughout malaria endemic countries and surveillance suffers from the lack of low-cost, high throughput genotyping methods. Molecular inversion probes (MIPs) allow for single-tube multiplexing of several thousand targets within the genome, sample barcoding and pooling of hundreds or thousands samples in a single sequencing run. We designed a MIP panel to target key insecticide resistance genes in Anopheles gambiae and An. arabiensis, including ace1, para, rdl, gste cluster genes, and several cyp6 genes, totalling 371 probes over 12 genes. Notably, MIP capture allowed us to tile 148 regions on para, covering nearly all 32 exons. Several rounds of optimization were performed using DNA from publicly available strains and cell lines, and sequencing was performed on both Illumina and Nanopore platforms. Initial analysis demonstrated high capture efficiency from magnetic bead extracted DNA from flash-frozen whole mosquitoes, as well as efficient capture from cDNA derived from dissected tissues. Variant calling with MIPtools software generated results concordant with expected strain genotypes. This panel bridges the gap between single gene dideoxy sequencing and intensive whole genome sequencing while overcoming the limitations of amplicon pools. Future work will apply this method to field-caught mosquitoes and is expected to dramatically reduce the cost of large-scale insecticide surveillance studies.