Bacteriophage‐Activated DNAzyme Hydrogels Combined with Machine Learning Enable Point‐of‐Use Colorimetric Detection of Escherichia coli

Abstract Developing cost‐effective, consumer‐accessible platforms for point‐of‐use environmental and clinical pathogen testing is a priority, to reduce reliance on laborious, time‐consuming culturing approaches. Unfortunately, a system offering ultrasensitive detection capabilities in a form that requires little auxiliary equipment or training has remained elusive. Here, a colorimetric DNAzyme‐crosslinked hydrogel sensor is presented. In the presence of a target pathogen, DNAzyme cleavage results in hydrogel dissolution, yielding the release of entrapped gold nanoparticles in a manner visible to the naked eye. Recognizing that Escherichia coli holds high relevance within both environmental and clinical environments, an E. coli‐responsive DNAzyme is incorporated into this platform. Through the optimization of the hydrogel polymerization process and the discovery of bacteriophage‐induced DNAzyme signal amplification, 10¹ CFU mL⁻¹ E. coli is detected within real‐world lake water samples. Subsequent pairing with an artificial intelligence model removed ambiguity in sensor readout, offering 96% true positive and 100% true negative accuracy. Finally, high sensor specificity and stability results supported clinical use, where 100% of urine samples collected from patients with E. coli urinary tract infections are accurately identified. No false positives are observed when testing healthy samples. Ultimately, this platform stands to significantly improve population health by substantially increasing pathogen testing accessibility.