Fig. 5
Design of DNA-based molecular diagnostic tools for detection of CYP6P4a and CYP6P4b resistance alleles and impact on resistance phenotype. A ARMS-PCR assay for genotyping the CYP6P4b-D284E marker. B LNA probe-based assay for genotyping the CYP6P4a-M220I marker. Association of the CYP6P4b-D284E and the CYP6P4a-M220I mutations with insecticide resistance phenotype. Distribution of CYP6P4b-D284E resistance marker among F3 FANGxGHANA hybrid An. funestus mosquitoes exposed to (C) permethrin for 30 min, D permethrin for 60 min, and (E) alphacypermethrin for 60 min. Distribution of CYP6P4a-M220I resistance marker among F3 FANGxGHANA hybrid An. funestus mosquitoes exposed to (F) permethrin for 30 min, G permethrin for 60 min, and (H) alphacypermethrin for 60 min. 284E, 220I (R) and D284, M220 (S) allele frequency distributions between alive and dead mosquitoes are shown in line plots. Combined impact of CYP6P4b-D284E and CYP6P4a-M220I mutations on pyrethroid insecticide resistance. Distribution of genotypes in FANGxGHANA hybrid An. funestus mosquitoes exposed to (J) permethrin for 60 min and (K) alphacypermethrin for 60 min with odds ratio calculations to compare the survival capacity of double homozygote-resistant mosquitoes after pyrethroid exposure with other genotype combinations