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Technologies that do not rely on growth cultures, nucleic acid amplification, step\u2010wise reagent addition, and complex sample processing are the key for meeting this need. Herein, multiple materials technologies are integrated for overcoming the obstacles in creating rapid and one\u2010pot bacterial sensing platforms. Liquid\u2010infused nanoelectrodes are developed for reducing nonspecific binding on the transducer surface; bacterium\u2010specific RNA\u2010cleaving DNAzymes are used for bacterial identification; and redox DNA barcodes embedded into DNAzymes are used for binding\u2010induced electrochemical signal transduction. The resultant single\u2010step and one\u2010pot assay demonstrates a limit\u2010of\u2010detection of 10<jats:sup>2<\/jats:sup> CFU\u00a0mL<jats:sup>\u22121<\/jats:sup>, with high specificity in identifying <jats:italic>Escherichia coli<\/jats:italic> amongst other Gram positive and negative bacteria including <jats:italic>Klebsiella pneumoniae<\/jats:italic>, <jats:italic>Staphylococcus aureus<\/jats:italic>, and <jats:italic>Bacillus subtilis<\/jats:italic>. Additionally, this assay is evaluated for analyzing 31 clinically obtained urine samples, demonstrating a clinical sensitivity of 100% and specify of 100%. When challenging this assay with nine clinical blood cultures, <jats:italic>E. coli<\/jats:italic>\u2010positive and <jats:italic>E. coli<\/jats:italic>\u2010negative samples can be distinguished with a probability of <jats:italic>p<\/jats:italic>\u00a0&lt;\u00a00.001.<\/jats:p>","DOI":"10.1002\/advs.202207223","type":"journal-article","created":{"date-parts":[[2023,4,23]],"date-time":"2023-04-23T22:41:29Z","timestamp":1682289689000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":28,"title":["Liquid NanoBiosensors Enable One\u2010Pot Electrochemical Detection of Bacteria in Complex Matrices"],"prefix":"10.1002","volume":"10","author":[{"given":"Sara M.","family":"Imani","sequence":"first","affiliation":[{"name":"School of Biomedical Engineering McMaster University  1280 Main Street West Hamilton ON L8S 4L7 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