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diagnostic platforms are desirable to surpass the deficiencies of conventional laboratory diagnostic methods for bacterial infections and to tackle the growing antimicrobial resistance crisis. In this study, a workflow was implemented, comprising the identification of new aptamers with high affinity for the ubiquitous surface protein A2 (UspA2) of the bacterial pathogen Moraxella catarrhalis and the development of an electrochemical biosensor functionalized with the best-performing aptamer as a bioreceptor to detect UspA2. After cell-systematic evolution of ligands by exponential enrichment (cell-SELEX) was performed, next-generation sequencing was used to sequence the final aptamer pool. The most frequent aptamer sequences were further evaluated using bioinformatic tools. The two most promising aptamer candidates, Apt1 and Apt1_RC (Apt1 reverse complement), had Kd values of 214.4 and 3.4 nM, respectively. Finally, a simple and label-free electrochemical biosensor was functionalized with Apt1_RC. The aptasensor surface modifications were confirmed by impedance spectroscopy and cyclic voltammetry. The ability to detect UspA2 was evaluated by square wave voltammetry, exhibiting a linear detection range of 4.0 \u00d7 104\u20137.0 \u00d7 107 CFU mL\u22121, a square correlation coefficient superior to 0.99 and a limit of detection of 4.0 \u00d7 104 CFU mL\u22121 at pH 5.0. The workflow described has the potential to be part of a sensitive PoC diagnostic platform to detect and quantify M. catarrhalis from biological samples.<\/jats:p>","DOI":"10.3390\/bioengineering10020178","type":"journal-article","created":{"date-parts":[[2023,2,1]],"date-time":"2023-02-01T02:06:44Z","timestamp":1675217204000},"page":"178","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Aptasensor for the Detection of Moraxella catarrhalis Adhesin UspA2"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5697-9292","authenticated-orcid":false,"given":"Maria G.","family":"Sande","sequence":"first","affiliation":[{"name":"CEB\u2014Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, 4710-057 Braga, Portugal"},{"name":"LABBELS\u2014Associate Laboratory, 4710-057 Braga, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0937-4047","authenticated-orcid":false,"given":"D\u00e9bora","family":"Ferreira","sequence":"additional","affiliation":[{"name":"CEB\u2014Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, 4710-057 Braga, Portugal"},{"name":"LABBELS\u2014Associate Laboratory, 4710-057 Braga, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3217-2320","authenticated-orcid":false,"given":"Joana L.","family":"Rodrigues","sequence":"additional","affiliation":[{"name":"CEB\u2014Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, 4710-057 Braga, Portugal"},{"name":"LABBELS\u2014Associate Laboratory, 4710-057 Braga, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6865-6044","authenticated-orcid":false,"given":"Lu\u00eds D. R.","family":"Melo","sequence":"additional","affiliation":[{"name":"CEB\u2014Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, 4710-057 Braga, Portugal"},{"name":"LABBELS\u2014Associate Laboratory, 4710-057 Braga, Portugal"}]},{"given":"Athanasios","family":"Saragliadis","sequence":"additional","affiliation":[{"name":"Section for Genetics and Evolutionary Biology, Department of Biosciences, University of Oslo, 0316 Oslo, Norway"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3150-6752","authenticated-orcid":false,"given":"Dirk","family":"Linke","sequence":"additional","affiliation":[{"name":"Section for Genetics and Evolutionary Biology, Department of Biosciences, University of Oslo, 0316 Oslo, Norway"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4237-8952","authenticated-orcid":false,"given":"Felismina T. 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