Summary of the technology
In the present study, we implement the isothermal rolling circle amplification (RCA) process to trigger a dendritic RCAinduced formation of amplifying DNAzyme catalytic labels. We describe the dendritic RCA-mediated formation of the Mg2+- dependent DNAzyme or of the hemin/G-quadruplex HRPmimicking DNAzyme units and the use of fluorescence, color, or chemiluminescence as readout signals for the different sensing platforms. Finally, by applying two different circular DNA templates, the multiplexed analysis of two different genes is demonstrated with the parallel detection of the Tay-Sachs genetic disorder mutant and the gene associated with the TP53 pathogen.
Project ID : 6-2014-2993
Description of the technology
The technology enables an affordable and easily deployable method for detection of water/soil contamination, and/or rapid detection of viruses in humans and animals.
The method employs the extension of the isothermal rolling circle amplification (RCA) process to a RCA-stimulated dendritic synthesis of catalytic nucleic acids (DNAzymes) sensing platform.
The method is based on the improvement of the RCA sensing methods by two elements:
- The introduction of a functional hairpin that regenerates the analyte and allows the dendritic branching of the RCA chains.
- The isothermal autonomous formation of DNAzymes in the RCA chains.
These catalytic labels enable simple detection of the chemical constituents of interest. By the tailoring of different circular DNA templates that lead to the formation of different RCA-branched DNAzyme reporter catalysts, the multiplexed analysis of different analytes is demonstrated.