Fig. 3

Dilution series of gametocytes tested by RT-PCR for 18S rRNA, Pf3D7_0630000 and pfs25. a RT-PCR was performed with hydrolysis probes on nucleic acids from enriched gametocyte cultures diluted into whole human blood across the range of parasite densities indicated in the figure (3 × 10⁷ gametocyte/mL blood to 3 gametocytes/mL blood). Samples for 18S rRNA and PF3D7_0630000 were not treated with DNase; samples for pfs25 RT-PCR were DNase treated as required for an unspliced target. DNase-treated 18S rRNA C_T values were comparable to non-DNase-treated C_Ts indicating that little to no 18S rRNA was lost in the DNase treatment (data not shown). Duplicates shown with Y-axis = ΔRn (change in fluorescence) from the Abbott m2000rt instrument. Labels indicate estimated parasite densities in parasites/mL (see Table 2 for log₁₀ values used in panel b. b Linear regression was performed using nominal log₁₀ transformed parasite densities and mean C_Ts (error bars 95% confidence intervals for available duplicate samples) to generate standard curves for the enriched gametocyte material