Simple, rapid, and accurate diagnosis is indispensable for broadercontrol measures, search for routes of infection, and understanding treatment status of emerging infectious diseases. Although PCR and LAMP are commonly used, in cases where the pathogen is an RNA virus, there are issues in terms of simplicity and speed, as multi-step operations including reverse transcription (RT) and many reagents, as well as time and expertise in optimizing primers and probes, are required. To solve these issues, we propose the artificial chaperone-enhanced MNAzyme (ACEzyme), which combines the functions of the multicomponent nucleic acid enzyme (MNAzyme) and cationic copolymers. PLL-g-Dex has artificial nucleic acid chaperone activity and can act as a molecular sensing system. ACEzyme resulted in an approximately 2,700-fold increase in MNAzyme activity. In this study, we showed that ACEzyme was also useful when working with RNA targets with robust high-order structures, and had high selectivity for single nucleotide mutation detection.
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