4.5 Article

Molecular Markers for Sensitive Detection of Plasmodium falciparum Asexual Stage Parasites and their Application in a Malaria Clinical Trial

Journal

AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE
Volume 97, Issue 1, Pages 188-198

Publisher

AMER SOC TROP MED & HYGIENE
DOI: 10.4269/ajtmh.16-0893

Keywords

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Funding

  1. Netherlands Organization for Scientific Research (Vidi fellowship
  2. NWO project) [016.158.306]
  3. Netherlands organization for international cooperation in higher education (Nuffic) [NFP-PhD. 14/150]
  4. Bill & Melinda Gates Foundation [AFIRM OPP1034789]
  5. European Research Council [ERC-2014-StG 639776]

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Plasmodium falciparum parasite life stages respond differently to antimalarial drugs. Sensitive stagespecific molecular assays may help to examine parasite dynamics at microscopically detectable and submicroscopic parasite densities in epidemiological and clinical studies. In this study, we compared the performance of skeleton-binding protein 1 (SBP1), ring-infected erythrocyte surface antigen, Hyp8, ring-exported protein 1 (REX1), and PHISTb mRNA for detecting ring-stage trophozoite-specific transcripts using quantitative reverse transcriptase polymerase chain reaction. Markers were tested on tightly synchronized in vitro parasites and clinical trial samples alongside established markers of parasite density (18S DNA and rRNA) and gametocyte density (Pfs25 mRNA). SBP1 was the most sensitive marker but showed low-level expression in mature gametocytes. Novel markers REX1 and PHISTb showed lower sensitivity but higher specificity for ring-stage trophozoites. Using in vivo clinical trial samples from gametocyte-negative patients, we observed evidence of persisting trophozoite transcripts for at least 14 days postinitiation of treatment. It is currently not clear if these transcripts represent viable parasites that may have implications for clinical treatment outcome or transmission potential.

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