Specific detection of topoisomerase i from the malaria causing P. falciparum parasite using isothermal Rolling Circle Amplification

We present a Rolling-Circle-Enhance-Enzyme-Activity-Detection (REEAD) system with potential use for future point-of-care diagnosis of malaria. In the developed setup, specific detection of malaria parasites in crude blood samples is facilitated by the conversion of single Plasmodium falciparum topoi...

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Bibliographic Details
Main Authors: Tesauro, Cinzia, Juul, Sissel, Arno, Barbara, Nielsen, Christine J.F., Fiorani, Paola, Frohlich, Rikke F., Andersen, Felicie F., Desideri, Alessandro, Stougaard, Magnus, Petersen, Eskild, Knudsen, Birgitta R.
Format: Conference Proceeding
Language:English
Subjects:
Biochemistry
Blood
Diseases
DNA
Educational institutions
Humans
Substrates
Online Access:Request full text
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Summary:We present a Rolling-Circle-Enhance-Enzyme-Activity-Detection (REEAD) system with potential use for future point-of-care diagnosis of malaria. In the developed setup, specific detection of malaria parasites in crude blood samples is facilitated by the conversion of single Plasmodium falciparum topoisomerase I (pfTopI) mediated cleavage-ligation events, happening within nanometer dimensions, to micrometer-sized products readily detectable at the single molecule level in a fluorescence microscope. In principle, REEAD requires no special equipment and the readout is adaptable to simple colorimetric detection systems. Moreover, with regard to detection limit the presented setup is likely to outcompete standard gold immuno-based diagnostics. Hence, we believe the presented assay forms the basis for a new generation of easy-to-use diagnostic tools suitable for the malaria epidemic areas in developing countries.
ISSN:1094-687X
1558-4615
2694-0604
DOI:10.1109/EMBC.2012.6346451