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

Abstract Plasmodium falciparum parasite life stages respond differently to antimalarial drugs. Sensitive stage-specific 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.
PMID
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Authors

Mayor MeshTerms
Keywords
Journal Title the american journal of tropical medicine and hygiene
Publication Year Start




PMID- 28719294
OWN - NLM
STAT- MEDLINE
DA  - 20170718
DCOM- 20170731
LR  - 20170731
IS  - 1476-1645 (Electronic)
IS  - 0002-9637 (Linking)
VI  - 97
IP  - 1
DP  - 2017 Jul
TI  - Molecular Markers for Sensitive Detection of Plasmodium falciparum Asexual Stage 
      Parasites and their Application in a Malaria Clinical Trial.
PG  - 188-198
LID - 10.4269/ajtmh.16-0893 [doi]
AB  - Plasmodium falciparum parasite life stages respond differently to antimalarial
      drugs. Sensitive stage-specific 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.
FAU - Tadesse, Fitsum G
AU  - Tadesse FG
AD  - Armauer Hansen Research Institute (AHRI), Addis Ababa, Ethiopia.
AD  - Medical Biotechnology Unit, Institute of Biotechnology, Addis Ababa University,
      Addis Ababa, Ethiopia.
AD  - Department of Medical Microbiology, Radboud University Medical Center, Nijmegen, 
      The Netherlands.
FAU - Lanke, Kjerstin
AU  - Lanke K
AD  - Department of Medical Microbiology, Radboud University Medical Center, Nijmegen, 
      The Netherlands.
FAU - Nebie, Issa
AU  - Nebie I
AD  - Department of Biomedical Sciences, Centre National de Recherche et de Formation
      sur le Paludisme, Ouagadougou,Burkina Faso.
FAU - Schildkraut, Jodie A
AU  - Schildkraut JA
AD  - Department of Medical Microbiology, Radboud University Medical Center, Nijmegen, 
      The Netherlands.
FAU - Goncalves, Bronner P
AU  - Goncalves BP
AD  - Department of Immunology and Infection, London School of Hygiene and Tropical
      Medicine, London, United Kingdom.
FAU - Tiono, Alfred B
AU  - Tiono AB
AD  - Department of Biomedical Sciences, Centre National de Recherche et de Formation
      sur le Paludisme, Ouagadougou,Burkina Faso.
FAU - Sauerwein, Robert
AU  - Sauerwein R
AD  - Department of Medical Microbiology, Radboud University Medical Center, Nijmegen, 
      The Netherlands.
FAU - Drakeley, Chris
AU  - Drakeley C
AD  - Department of Immunology and Infection, London School of Hygiene and Tropical
      Medicine, London, United Kingdom.
FAU - Bousema, Teun
AU  - Bousema T
AD  - Department of Medical Microbiology, Radboud University Medical Center, Nijmegen, 
      The Netherlands.
AD  - Department of Immunology and Infection, London School of Hygiene and Tropical
      Medicine, London, United Kingdom.
FAU - Rijpma, Sanna R
AU  - Rijpma SR
AD  - Department of Laboratory Medicine, Radboud University Medical Center, Nijmegen,
      The Netherlands.
AD  - Department of Medical Microbiology, Radboud University Medical Center, Nijmegen, 
      The Netherlands.
LA  - eng
PT  - Journal Article
PT  - Randomized Controlled Trial
PL  - United States
TA  - Am J Trop Med Hyg
JT  - The American journal of tropical medicine and hygiene
JID - 0370507
RN  - 0 (Antimalarials)
RN  - 0 (Artemisinins)
RN  - 0 (Biomarkers)
RN  - 0 (DNA, Protozoan)
RN  - 0 (Drug Combinations)
RN  - 0 (Ethanolamines)
RN  - 0 (Fluorenes)
RN  - 0 (RNA, Protozoan)
RN  - 0 (RNA, Ribosomal, 18S)
RN  - 0 (artemether-lumefantrine combination)
RN  - MVR3634GX1 (Primaquine)
SB  - AIM
SB  - IM
MH  - Antimalarials/therapeutic use
MH  - Artemisinins/administration & dosage/therapeutic use
MH  - Biomarkers/blood
MH  - Burkina Faso/epidemiology
MH  - DNA, Protozoan/genetics
MH  - Double-Blind Method
MH  - Drug Combinations
MH  - Ethanolamines/administration & dosage/therapeutic use
MH  - Fluorenes/administration & dosage/therapeutic use
MH  - Humans
MH  - Malaria, Falciparum/*blood/drug therapy/epidemiology/*parasitology
MH  - Plasmodium falciparum/genetics/*isolation & purification
MH  - Primaquine/administration & dosage/therapeutic use
MH  - RNA, Protozoan/genetics
MH  - RNA, Ribosomal, 18S/genetics/isolation & purification
MH  - Trophozoites/metabolism
PMC - PMC5508903
EDAT- 2017/07/19 06:00
MHDA- 2017/08/02 06:00
CRDT- 2017/07/19 06:00
AID - 10.4269/ajtmh.16-0893 [doi]
PST - ppublish
SO  - Am J Trop Med Hyg. 2017 Jul;97(1):188-198. doi: 10.4269/ajtmh.16-0893.