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 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.

Citation

Cinzia Tesauro, Sissel Juul, Barbara Arnò, Christine J F Nielsen, Paola Fiorani, Rikke F Frøhlich, Felicie F Andersen, Alessandro Desideri, Magnus Stougaard, Eskild Petersen, Birgitta R Knudsen. Specific detection of topoisomerase I from the malaria causing P. falciparum parasite using isothermal rolling circle amplification. Conference proceedings : ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference. 2012;2012:2416-9

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PMID: 23366412

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