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To develop a new electrochemical DNA biosensor for determination of Klebsiella pneumoniae carbapenemase, a highly sensitive and selective electrochemical biosensor for DNA detection was constructed based on a glassy carbon electrode (GCE) modified with gold nanoparticles (Au-nano). The Au-nano/GCE was characterized by scanning electromicroscopy, cyclic voltammetry, and electrochemical impedance spectroscopy. The hybridization detection was measured by differential pulse voltammetry using methylene blue as the hybridization indicator. The dynamic range of detection of the sensor for the target DNA sequences was from 1 × 10–11 to 1 × 10–8 M, with an LOD of 1 × 10–12 M. The DNA biosensor had excellent specificity for distinguishing complementary DNA sequence in the presence of non-complementary and mismatched DNA sequence. The Au-nano/GCE showed significant improvement in electrochemical characteristics, and this biosensor was successfully applied for determination of K. pneumoniae.

A label-free DNA hybridization electrochemical sensor for the detection of Klebsiella pneumoniae was developed, which could be helpful in the diagnosis of bacterial infections. Indole-5-carboxylic acid (ICA) and graphene oxide (GO) were electrodeposited on a glassy carbon electrode, and the resulting reduced GO (rGO)-ICA hybrid film served as a platform for immobilizing oligonucleotides on a single-stranded DNA (ssDNA) sequence. The conditions were optimized, with excellent electrochemical performance. A significant change was observed after hybridization of ssDNA with the target probe under optimum conditions. Hybridization with complementary, noncomplementary, one-base mismatched, and three-base mismatched DNA targets was studied effectively by differential pulse voltammetry. The proposed strategy could detect target DNA down to 3 × 10-11 M, with a linear range from 1 × 10-6 M to 1 × 10-10 M, showing high sensitivity. This electrochemical method is simple, free from indicator, and shows good selectivity. Hence, electrochemical biosensors are successfully demonstrated for the detection of K. pneumoniae.