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Sensitive and accurate quantification of human malaria parasites using droplet digital PCR (ddPCR).

  • Koepfli C Walter &Eliza Hall Institute, Parkville, Australia.
  • Nguitragool W Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
  • Hofmann NE Swiss Tropical and Public Health Institute, Basel, Switzerland.
  • Robinson LJ Walter &Eliza Hall Institute, Parkville, Australia.
  • Ome-Kaius M Papua New Guinea Institute of Medical Research, Madang, Papua New Guinea.
  • Sattabongkot J Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
  • Felger I Swiss Tropical and Public Health Institute, Basel, Switzerland.
  • Mueller I Walter &Eliza Hall Institute, Parkville, Australia.
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  • 2016-12-17
Published in:
  • Scientific reports. - 2016
DNA, Protozoan
Humans
Malaria
Microscopy
Plasmodium falciparum
Plasmodium vivax
RNA, Ribosomal, 18S
Real-Time Polymerase Chain Reaction
English Accurate quantification of parasite density in the human host is essential for understanding the biology and pathology of malaria. Semi-quantitative molecular methods are widely applied, but the need for an external standard curve makes it difficult to compare parasite density estimates across studies. Droplet digital PCR (ddPCR) allows direct quantification without the need for a standard curve. ddPCR was used to diagnose and quantify P. falciparum and P. vivax in clinical patients as well as in asymptomatic samples. ddPCR yielded highly reproducible measurements across the range of parasite densities observed in humans, and showed higher sensitivity than qPCR to diagnose P. falciparum, and equal sensitivity for P. vivax. Correspondence in quantification was very high (>0.95) between qPCR and ddPCR. Quantification between technical replicates by ddPCR differed 1.5-1.7-fold, compared to 2.4-6.2-fold by qPCR. ddPCR facilitates parasite quantification for studies where absolute densities are required, and will increase comparability of results reported from different laboratories.
Language
  • English
Open access status
gold
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Persistent URL
https://sonar.ch/global/documents/56552
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