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The prepared biosensor showed satisfactory selectivity, reproducibility, and stability towards miRNA-21 detection. The biosensor was feasible for accurate and quantitative detection of miRNA-21 in normal human serum samples with RSD below 5.86%, which showed a great potential in clinical analysis and disease diagnosis.Escherichia coli and Enterococcus faecalis are two of the most prevalent uro-pathogens and are difficult to treat as they acquire multidrug-resistant traits. In this study, the main objective was to develop biocompatible co