Abstract

The detection of pathogens is critical for preventing and controlling health hazards across clinical, environmental, and food safety sectors. Functional nucleic acids (FNAs), such as aptamers and DNAzymes, have emerged as versatile molecular tools for pathogen detection due to their high specificity and affinity. This review focuses on the in vitro selection of FNAs for pathogens, with emphasis on the selection of aptamers for specific biomarkers and intact pathogens, including bacteria and viruses. Additionally, the selection of DNAzymes for bacterial detection is discussed. The integration of these FNAs into colorimetric biosensors has enabled the development of simple, cost‐effective diagnostic platforms. Both non‐catalytic and catalytic colorimetric biosensors are explored, including those based on gold nanoparticles (AuNPs), polydiacetylenes (PDAs), protein enzymes, G‐quadruplexes, and nanozymes. These biosensors offer visible detection through color changes, making them ideal for point‐of‐care diagnostics. The review concludes by highlighting current challenges and future perspectives for advancing FNA‐based colorimetric biosensing technologies for pathogen detection.

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