Allele-Specific PCR for TEP1 Genotyping

Two TEP1 allele-specific PCR assays were employed to genotype the variants previously described in An. gambiae populations [4 (link), 12 (link)]. The first assay [4 (link)] amplifies a 428 bp fragment for the susceptible allele (TEP1s) and 349 bp fragment for the resistant allele (TEP1r). The genotypes of TEP1r were amplified using the second assay [12 (link)], which targets 510 bp fragment for TEP1r^A and 155 bp fragment for TEP1r^B. The wild type (TEP1) (646 bp) was also amplified from the second assay. PCR products were analysed using QIAxcel capillary electrophoresis to identify the different fragments. Only fragments that were positive from both PCR assays were finally analysed. A total of 1400 mosquitoes were genotyped (2009 = 335; 2016 = 525; 2019 = 540).