A Molecular Assay to Quantify Male and Female Plasmodium falciparum Gametocytes: Results From 2 Randomized Controlled Trials Using Primaquine for Gametocyte Clearance

J Infect Dis. 2017 Aug 15;216(4):457-467. doi: 10.1093/infdis/jix237.

Abstract

Background: Single low-dose primaquine (PQ) reduces Plasmodium falciparum infectivity before it impacts gametocyte density. Here, we examined the effect of PQ on gametocyte sex ratio as a possible explanation for this early sterilizing effect.

Methods: Quantitative reverse-transcription polymerase chain reaction assays were developed to quantify female gametocytes (targeting Pfs25 messenger RNA [mRNA]) and male gametocytes (targeting Pf3D7_1469900 mRNA) in 2 randomized trials in Kenya and Mali, comparing dihydroartemisinin-piperaquine (DP) alone to DP with PQ. Gametocyte sex ratio was examined in relation to time since treatment and infectivity to mosquitoes.

Results: In Kenya, the median proportion of male gametocytes was 0.33 at baseline. Seven days after treatment, gametocyte density was significantly reduced in the DP-PQ arm relative to the DP arm (females: 0.05% [interquartile range {IQR}, 0.0-0.7%] of baseline; males: 3.4% [IQR, 0.4%-32.9%] of baseline; P < .001). Twenty-four hours after treatment, gametocyte sex ratio became male-biased and was not significantly different between the DP and DP-PQ groups. In Mali, there was no significant difference in sex ratio between the DP and DP-PQ groups (>0.125 mg/kg) 48 hours after treatment, and gametocyte sex ratio was not associated with mosquito infection rates.

Conclusions: The early sterilizing effects of PQ may not be explained by the preferential clearance of male gametocytes and may be due to an effect on gametocyte fitness.

Keywords: artemisinin; dihydroartemisinin; piperaquine; primaquine; sex ratio.

Publication types

  • Randomized Controlled Trial

MeSH terms

  • Adolescent
  • Antimalarials / therapeutic use*
  • Artemisinins / therapeutic use
  • Child
  • Child, Preschool
  • Dose-Response Relationship, Drug
  • Female
  • Germ Cells / drug effects*
  • Humans
  • Kenya
  • Male
  • Mali
  • Plasmodium falciparum
  • Primaquine / therapeutic use*
  • Protozoan Proteins / genetics*
  • Protozoan Proteins / metabolism
  • Quinolines / therapeutic use
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Sample Size

Substances

  • Antimalarials
  • Artemisinins
  • Pfs25 protein, Plasmodium falciparum
  • Protozoan Proteins
  • Quinolines
  • RNA, Messenger
  • artenimol
  • piperaquine
  • Primaquine