A Programmable DNAzyme for the Sensitive Detection of Nucleic Acids

Area of Science:

• Biomolecular Engineering
• Molecular Diagnostics
• Biosensor Technology

Background:

• Nucleic acids in biofluids are promising disease biomarkers.
• Current detection assays lack the sensitivity for widespread clinical use.
• Sensitive and robust detection methods are crucial for molecular diagnostics.

Purpose of the Study:

• To develop a sensitive nucleic acid detection assay named SPOT.
• To enable early and accurate molecular diagnostics of various diseases.
• To create a versatile platform for detecting low-abundance nucleic acids.

Main Methods:

• Designed a catalytic DNAzyme with endonuclease capability.
• Integrated the DNAzyme into a one-stranded allosteric biosensor (SPOT).
• Validated SPOT using serum microRNAs (miRNAs) and SARS-CoV-2 RNA from clinical samples.

Main Results:

• SPOT demonstrates high sensitivity, specificity, and robustness in detecting nucleic acids.
• Successfully detected serum miRNAs for breast, gastric, and prostate cancer diagnostics.
• Achieved potent detection of SARS-CoV-2 RNA from clinical swabs.
• SPOT is compatible with point-of-care testing (POCT) platforms like lateral flow assays.

Conclusions:

• SPOT is a highly sensitive, convenient, and cost-effective assay for nucleic acid detection.
• Enables molecular diagnostics for various cancers and infectious diseases.
• Offers a robust platform for point-of-care testing and clinical applications.