Open AccessArticle
Refined Method for Droplet Microfluidics-Enabled Detection of Plasmodium falciparum Encoded Topoisomerase I in Blood from Malaria Patients
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Marianne Smedegaard Hede, Patricia Nkem Okorie, Signe Kirk Fruekilde, Søren Fjelstrup, Jonas Thomsen, Oskar Franch, Cinzia Tesauro, Magnus Tobias Bugge, Mette Christiansen, Stéphane Picot, Felix Lötsch, Ghyslain Mombo-Ngoma, Johannes Mischlinger, Ayôla A. Adegnika, Finn Skou Pedersen, Yi-Ping Ho, Eskild Petersen, Magnus Stougaard, Michael Ramharter and Birgitta Ruth Knudsen
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Abstract
Rapid and reliable diagnosis is essential in the fight against malaria, which remains one of the most deadly infectious diseases in the world. In the present study we take advantage of a droplet microfluidics platform combined with a novel and user-friendly biosensor for
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Rapid and reliable diagnosis is essential in the fight against malaria, which remains one of the most deadly infectious diseases in the world. In the present study we take advantage of a droplet microfluidics platform combined with a novel and user-friendly biosensor for revealing the main malaria-causing agent, the
Plasmodium falciparum (P. falciparum) parasite. Detection of the parasite is achieved through detection of the activity of a parasite-produced DNA-modifying enzyme, topoisomerase I (pfTopoI), in the blood from malaria patients. The assay presented has three steps: (1) droplet microfluidics-enabled extraction of active pfTopoI from a patient blood sample; (2) pfTopoI-mediated modification of a specialized DNA biosensor; (3) readout. The setup is quantitative and specific for the detection of
Plasmodium topoisomerase I. The procedure is a considerable improvement of the previously published Rolling Circle Enhanced Enzyme Activity Detection (REEAD) due to the advantages of involving no signal amplification steps combined with a user-friendly readout. In combination these alterations represent an important step towards exploiting enzyme activity detection in point-of-care diagnostics of malaria.
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