Journal article

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

Cristian Koepfli, Wang Nguitragool, Natalie E Hofmann, Leanne J Robinson, Maria Ome-Kaius, Jetsumon Sattabongkot, Ingrid Felger, Ivo Mueller

Scientific Reports | NATURE PUBLISHING GROUP | Published : 2016

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..

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Grants

Awarded by NHMRC


Awarded by Swiss National Science Foundation


Awarded by NIH International Centers of Excellence in Malaria Research grant


Awarded by SNF Early Postdoc Mobility Fellowship


Awarded by NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES


Funding Acknowledgements

We would like to thank staff of the Papua New Guinea Institute of Medical Research and Mahidol Vivax Research Unit for help with collection of samples, and all patients and communities for their consent to participate in the field studies. We thank Nattawan Rachaphaew, Lina Lorry, Samuel Maripal and Charles Kongs for parasite counts by light microscopy, and Annie Yang for providing P. falciparum culture. This study was supported by the TransEPI consortium funded by the Bill & Melinda Gates Foundation, and an NHMRC project grant (#1021455), Swiss National Science Foundation grant (310030_134889), and NIH International Centers of Excellence in Malaria Research grant (U19 AI089686). C.K. was supported by an SNF Early Postdoc Mobility Fellowship (#P2BSP3_151880). This work was made possible through Victorian State Government Operational Infrastructure Support and Australian Government NHMRC IRIISS.