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.