Modular DNAzymes-Hydrogel Membrane Carriers for Highly Sensitive Isothermal Cross-Cascade Detection of Pathogenic Bacteria Nucleic Acids

Abstract

The increasing prevalence of antimicrobial resistance has called for improved diagnostic testing of pathogenic bacteria. However, the development of rapid, cost-effective, and easy-to-use tests for bacterial infections remains a constant challenge. Here, we report a class of modular hydrogel membrane carriers incorporated with composite DNAzymes, which enable rapid and highly sensitive detection of pathogenic bacteria gene target analytes. We apply free radical polymerization to incorporate composite DNAzymes, consisting of an RNA substrate component and a DNAzyme component (e.g., 10−23 or 8−17 DNAzymes), into polyethylene glycol diacrylate polymer networks. Initiated by a nucleic acid target acting as an assembly facilitator, multicomponent DNAzymes are combined to cleave the RNA substrate component in the hydrogel carriers, which releases the DNAzyme component to cleave RNA reporter probes to generate fluorescence. We modulate the morphology, composition, and microporous structures of the DNAzyme carriers to achieve quantitative assay performance. We demonstrate a rapid and highsensitivity detection of C. trachomatis gene target analytes as low as 50 fM in a short assay time of 25 min. The work represents a crucial step forward in the development of a generic, isothermal, and protein enzyme-free pathogenic bacteria testing platform technology.

Background-Free and Reversible Upconversion Hydrogel Sensing Platform for Visual Monitoring of Sulfite

“…In contrast, porous polymer network hydrogels have become one of the important carriers of fluorescent probes due to their semiwet structure, good mechanical stability, and adjustable size. 27,28 The fluorescent probe can be uniformly loaded into the hydrogel network during the preparation of hydrogel. At the same time, the hydrogel provides a porous semiwet structure, which maintains the same high selectivity and sensitivity detection compared to that in the liquid environ- 31,32 Hence, designing a UCNP-based luminescent hydrogel sensor for the detection of SO 2 derivatives is an effective strategy to avoid the autofluorescence generated by the hydrogel matrix.…”