Pseudomonas aeruginosa, an opportunistic Gram-negative bacterium, is one of the main sources of infections in healthcare environments, making its detection very important. N-3-oxo-dodecanoyl L-homoserine lactone (3-O-C₁₂-HSL) is a characteristic molecule of quorum sensing—a form of cell-to-cell communication between bacteria—in P. aeruginosa. Its detection can allow the determination of the bacterial population. In this study, the development of the first electrochemical aptasensor for the detection of 3-O-C₁₂-HSL is reported. A carbon-based screen-printed electrode modified with gold nanoparticles proved to be the best platform for the aptasensor. Each step in the fabrication of the aptasensor (i.e., gold nanoparticles’ deposition, aptamer immobilization, incubation with the analyte) was optimized and characterized using cyclic voltammetry, differential pulse voltammetry, and electrochemical impedance spectroscopy. Different redox probes in solution were evaluated, the best results being obtained in the presence of [Fe(CN)₆]⁴⁻/[Fe(CN)₆]³⁻. The binding affinity of 106.7 nM for the immobilized thiol-terminated aptamer was determined using surface plasmon resonance. The quantification of 3-O-C₁₂-HSL was performed by using the electrochemical signal of the redox probe before and after incubation with the analyte. The aptasensor exhibited a logarithmic range from 0.5 to 30 µM, with a limit of detection of 145 ng mL⁻¹ (0.5 µM). The aptasensor was successfully applied for the analysis of real samples (e.g., spiked urine samples, spiked microbiological growth media, and microbiological cultures).

The rapid diagnosis of Pseudomonas aeruginosa infection is very important because this bacterium is one of the main sources of healthcare-associated infections. Pseudomonas quinolone signal (PQS) is a specific molecule for quorum sensing (QS) in P. aeruginosa, a form of cell-to-cell bacterial communication and its levels can allow the determination of the bacterial population. In this study, the development of the first electrochemical detection of PQS using screen-printed electrodes modified with carbon nanotubes (CNT-SPE) is reported. The electrochemical fingerprint of PQS was determined using different electrode materials and screen-printed electrodes modified with different nanomaterials. The optimization of the method in terms of electrolyte, pH, and electrochemical technique was achieved. The quantification of PQS was performed using one of the anodic peaks in the electrochemical fingerprint of the PQS on the CNT-SPE. The sensor exhibited a linear range from 0.1 to 15 µM, with a limit of detection of 50 nM. The sensor allowed the selective detection of PQS, with low interference from other QS molecules. The sensor was successfully applied to analysis of real samples (spiked urine and human serum samples, spiked microbiological growth media, and microbiological cultures).