Abstract
Multi-component nucleotide acid enzymes (MNAzymes) derived from RNase-mimic DNAzymes have potential as simple and accurate DNA detectors. To enhance the MNAzyme activity under multiple-turnover conditions, a cationic comb-type copolymer, PLL-g-Dex, that facilitates hybridization and strand exchange reactions of DNA was utilized. The copolymer increased the MNAzyme reaction rate by 200 times, allowing target DNA detection at picomolar concentrations at physiological temperature.
Highlights
The emergence of life-threatening infections such as severe acute respiratory syndrome (SARS) and viral haemorrhagic fevers caused by the Ebola and Marburg viruses highlights the urgent need for efficient infection control practices
Isothermal amplification methods,[1,2] such as rolling circle amplification (RCA),[3,4] loop-mediated isothermal amplification (LAMP),[5] and helicase-dependent amplification (HAD),[6] to detect pathogen genetic material or disease markers have advantages over current PCR-based methods because of their simple protocols and because they could be conducted at ambient temperature
In the presence of PLL-gDex, the sensitivity was enhanced approximately 5 times at an Multi-component nucleic acid enzyme (MNAzyme) concentration of 2 nM (Fig. 3B), indicating that the copolymer significantly improved the MNAzyme reactivity under multiple-turnover conditions. This suggests that the use of the copolymer could enable use of the MNAzyme assay for detection of short nucleotide targets such as miRNA and siRNA
Summary
The emergence of life-threatening infections such as severe acute respiratory syndrome (SARS) and viral haemorrhagic fevers caused by the Ebola and Marburg viruses highlights the urgent need for efficient infection control practices. Isothermal amplification methods,[1,2] such as rolling circle amplification (RCA),[3,4] loop-mediated isothermal amplification (LAMP),[5] and helicase-dependent amplification (HAD),[6] to detect pathogen genetic material or disease markers have advantages over current PCR-based methods because of their simple protocols and because they could be conducted at ambient temperature. These isothermal amplification methods still have complicated setups and require multiple enzymes and/or primers. The MNAzyme integrated with various detection methods has been widely investigated.[11,12,13,14,15,16]
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