Plasmodium infection in relation to the 4.3kb Structural variant.
Genotyping results for Obout 2016 samples revealed the presence of the three genotypes for the 4.3kb SV. Also, previous data had shown that mosquitoes collected in Obout in May 2016 had high Plasmodiuminfection rates. The Plasmodium infection rate was 38.7% (72/186), 79.2% of which was P. falciparum , 12.5% ovale-vivax-malariae OVM + and 8.3% mix infection (Nkemngo et al., 2022). Screening for Plasmodium infection using TaqMan assay was done on 186 whole mosquito specimens from Obout. The real-time PCR MX 3005 (Agilent, Santa Clara, CA, USA) system was used for the amplification (Bass et al., 2008). Briefly, 2 μL of gDNA for each sample was used as a template in a 3-step program with a pre-denaturation at 95 ◦C for 10 mins, followed by 40 cycles of 15 sec at 95 ◦C and 1 min at 60 ◦C. The primers (Falcip+: TCT GAA TAC GAA TGT C, OVM+: CTG AAT ACA AAT GCC, Plas-F: GCT TAG TTA CGA TTA ATA GGA GTA GCT TG, Plas R: GAA AAT CTA AGA ATT TCA CCT CTG ACA) were used together with two probes tagged with fluorophores: FAM to detect Plasmodium falciparum , and HEX to detect Plasmodium ovale , Plasmodium vivax, and P. malariae. P. falciparum samples and a mix of P. ovale , P. vivax, and P. malariae were used as positive controls. A sub-set of positive samples was subjected to Nested PCR to confirm and discriminate the species detected by TaqMan based on the protocol of (Snounou et al., 1993). Only the P. falciparum positives and non-infected were used to investigate the Plasmodium infection rates in mosquitoes with different genotypes for the 4.3kb SV. In total 79 mosquitoes were analysed.