Journal
ACS NANO
Volume 6, Issue 12, Pages 10676-10683Publisher
AMER CHEMICAL SOC
DOI: 10.1021/nn3038594
Keywords
droplet microfluidics; malaria; diagnosis; enzyme activity detection; rolling-circle amplification; lab-on-a-chip
Categories
Funding
- NIH [HL89764]
- NSF [EEC-0425626]
- Danish Research Councils [11-116325/FTP, 11-105736/FSS]
- Karen Elise Jensen's Foundation
- Dagmar Marshall's Foundation
- Dir. Einar Hansen og Vera Hansen's Foundation
- Harboe Foundation
- Augustinus Foundation
- Louis Hansen's Foundation
- Horslev Foundation
- Fabrikant Einar Willumsen's Foundation
- Kobmand Sven Hansen & hustru Ina Hansen's Foundation
- Dir. Emil Hertz & hustru Inger Hertz' Foundation
- Civilingenior Frode Nygaard's Foundation
- Kong Christian den Tiende's Foundation
- Gangsted Foundation
- KU's Foundation for Cancer Research
- Ludvig og Franciska Andersen's Foundation
- Arvid Nilsson's Foundation
- Frimodt-Heineke's Foundation
- Aase and Ejnar Danielsens Foundation
- Minister Erna Hamiltons Foundation for Science and Art
- Apolodoro Plausonius Foundation
- Italian Association for Cancer Research (AIRC)
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We present an attractive new system for the specific and sensitive detection of the malaria-causing Plasmodium parasites. The system relies on isothermal conversion of single DNA cleavage ligation events catalyzed specifically by the Plasmodium enzyme topoisomerase I to micrometer-sized products detectable at the single-molecule level. Combined with a droplet microfluidics lab-on-a-chip platform, this design allowed for sensitive, specific, and quantitative detection of all human-malaria-causing Plasmodium species in single drops of unprocessed blood with a detection limit of less than one parasite/mu L. Moreover, the setup allowed for detection of Plasmodium parasites in noninvasive saliva samples from infected patients. During recent years malaria transmission has declined worldwide, and with this the number of patients with low-parasite density has increased. Consequently, the need for accurate detection of even a few parasites is becoming increasingly important for the continued combat against the disease. We believe that the presented droplet microfluidics platform, which has a high potential for adaptation to point-of-care setups suitable for low-resource settings, may contribute significantly to meet this demand. Moreover, potential future adaptation of the presented setup for the detection of other microorganisms may form the basis for the development of a more generic platform for diagnosis, fresh water or food quality control, or other purposes within applied or basic science.
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