P. ovale spp. isolates were selected from the FNMRC database based on parasite densities and the countries where the patients got contaminated.
Genomic DNA was extracted from 200 μL of whole blood using MagNA Pure automaton (Roche diagnostics, USA) and eluted in 100 μL. P. ovale spp. mono-infection was confirmed with the species-specific quantitative PCR (qPCR) Plasmodium typage kit (Bio-Evolution, France) targeting the 18s rRNA for P. ovale spp. and the human beta actin gene to evaluate human DNA contamination. The reaction was carried out on a ViiA 7 thermocycler (Applied Biosystems). One positive and one negative control were included in each run. ΔCt (cycle threshold) was defined as the difference between the Ct of P. ovale spp. and the human Ct. We performed in-house qPCR high resolution melting (HRM) to differentiate P. ovale wallikeri from P. ovale curtisi (42 (link)).
Genomic DNA was extracted from 200 μL of whole blood using MagNA Pure automaton (Roche diagnostics, USA) and eluted in 100 μL. P. ovale spp. mono-infection was confirmed with the species-specific quantitative PCR (qPCR) Plasmodium typage kit (Bio-Evolution, France) targeting the 18s rRNA for P. ovale spp. and the human beta actin gene to evaluate human DNA contamination. The reaction was carried out on a ViiA 7 thermocycler (Applied Biosystems). One positive and one negative control were included in each run. ΔCt (cycle threshold) was defined as the difference between the Ct of P. ovale spp. and the human Ct. We performed in-house qPCR high resolution melting (HRM) to differentiate P. ovale wallikeri from P. ovale curtisi (42 (link)).