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Micromachines 2015, 6(10), 1505-1513; doi:10.3390/mi6101432

Refined Method for Droplet Microfluidics-Enabled Detection of Plasmodium falciparum Encoded Topoisomerase I in Blood from Malaria Patients

1
Zymonostics, 8000 Aarhus C, Denmark
2
Institute for Advanced Medical Research and Training, College of Medicine, University of Ibadan, 200001 Ibadan, Nigeria
3
Department of Molecular Biology and Genetics, University of Aarhus, 8000 Aarhus C, Denmark
4
NANO, University of Aarhus, 8000 Aarhus C, Denmark
5
Department of Clinical Immunology, Aarhus University Hospital, 8200 Aarhus N, Denmark
6
Malaria Research Unit, SMITh, ICBMS, UMR 5246 CNRS-INSA-CPE-UCBL1 & Institutde Parasitologie et Mycologie Médicale, Hospices Civils de Lyon, 69373 Lyon Cedex 08, France
7
Centre de Recherches Médicales de Lambaréné, Lambaréné, Gabon
8
Institut für Tropenmedizin, Universität Tübingen, 72074 Tübingen, Germany
9
Department of Infectious Diseases, Aarhus University Hospital, 8200 Aarhus N, Denmark
10
Department of Clinical Medicine, University of Aarhus, 8000 Aarhus C, Denmark
11
Department of Medicine I, Division of Infectious Diseases and Tropical Medicine, Medical University of Vienna, 1090 Vienna, Austria
*
Author to whom correspondence should be addressed.
Academic Editors: Andrew deMello and Xavier Casadevall i Solvas
Received: 17 August 2015 / Revised: 23 September 2015 / Accepted: 29 September 2015 / Published: 5 October 2015
(This article belongs to the Special Issue Droplet Microfluidics: Techniques and Technologies)
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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 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. View Full-Text
Keywords: malaria; point-of-care; droplet microfluidics; diagnostics; topoisomerase malaria; point-of-care; droplet microfluidics; diagnostics; topoisomerase
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Hede, M.S.; Okorie, P.N.; Fruekilde, S.K.; Fjelstrup, S.; Thomsen, J.; Franch, O.; Tesauro, C.; Bugge, M.T.; Christiansen, M.; Picot, S.; Lötsch, F.; Mombo-Ngoma, G.; Mischlinger, J.; Adegnika, A.A.; Pedersen, F.S.; Ho, Y.-P.; Petersen, E.; Stougaard, M.; Ramharter, M.; Knudsen, B.R. Refined Method for Droplet Microfluidics-Enabled Detection of Plasmodium falciparum Encoded Topoisomerase I in Blood from Malaria Patients. Micromachines 2015, 6, 1505-1513.

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