CRISPR/Cas-powered "sweet" DNA nano-sponge for highly sensitive detection of pathogen nucleic acid with portable personal glucometer

The strategy of coupling clustered regularly interspaced short palindromic repeats (CRISPR) system and personal glucometer holds significant promise for point-of-care pathogen nucleic acid testing. However, prevailing strategies that rely on the synthesis of nucleic acid-protein conjugates, which is inefficient and cumbersome. Herein, we present an amylase-encapsulated DNA nano-sponge designed to respond to the activated CRISPR system to produce glucose signal output. Owing to a high-affinity crosslinker and single-stranded DNA scaffolding, the proposed DNA nano-sponge ensures efficient amylase encapsulation and responsive release. By coupling with rapid nucleic acid amplification methods, the proposed detection platform was applied to specifically detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and influenza A virus, with a limit of detection as low as 101 copies of target per test. The accurate detection of real-world samples further demonstrated the great potential of the proposed platform for clinical applications and public health surveillance.