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)).
Molecular Detection of Plasmodium ovale Subspecies
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)).
Corresponding Organization : Institut de Recherche pour le Développement
Variable analysis
- Parasite densities
- Countries where patients got contaminated
- Confirmation of P. ovale spp. mono-infection using species-specific qPCR
- Differentiation of P. ovale wallikeri from P. ovale curtisi using in-house qPCR high resolution melting (HRM)
- 200 μL of whole blood used for genomic DNA extraction
- Elution volume of 100 μL after DNA extraction
- Inclusion of one positive and one negative control in each qPCR run
- Positive control
- Negative control
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