Sensitive and accurate quantification of human malaria parasites using droplet digital PCR (ddPCR)
Journal Title
Scientific Reports
Publication Type
Journal Article
Abstract
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.
Publisher
Springer Nature
WEHI Research Division(s)
Population Health And Immunity
PubMed ID
27982132
Publisher's Version
https://doi.org/10.1038/srep39183
Open Access at Publisher's Site
http://www.nature.com/articles/srep39183
Rights Notice
Refer to copyright notice on published article.


Creation Date: 2017-05-26 03:48:51
Last Modified: 2017-05-26 04:02:28
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