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.
Department of Biology and Center of Biostatistics and Bioinformatics & NAST Nanoscience & Nanotechnology & Innovative Instrumentation, University of Rome Tor Vergata, Rome, Italy.
Cinzia
C
Tesauro
Sissel
S
Juul
Barbara
B
Arnò
Christine J F
CJ
Nielsen
Paola
P
Fiorani
Rikke F
RF
Frøhlich
Felicie F
FF
Andersen
Alessandro
A
Desideri
Magnus
M
Stougaard
Eskild
E
Petersen
Birgitta R
BR
Knudsen
D015374:Biosensing Techniques / Q000379:methods*; D004264:DNA Topoisomerases, Type I / Q000097:blood* / Q000235:genetics*; D006801:Humans; D008288:Malaria / Q000175:diagnosis* / Q000469:parasitology*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010963:Plasmodium falciparum / Q000201:enzymology* / Q000302:isolation & purification
D004264:DNA Topoisomerases, Type I