Abstract
Hospital environments are recognized as significant reservoirs of nosocomial pathogens, contributing to the onset of healthcare-associated infections (HAIs). Timely microbiological monitoring is essential to mitigate infection risks. However, gold-standard methods based on culture and biochemical techniques are time-consuming and may underestimate microbial contamination, potentially delaying interventions. This study proposes a novel approach for surface monitoring using loop-mediated isothermal amplification (LAMP) for the rapid detection of key nosocomial pathogens, such as Pseudomonas aeruginosa, Staphylococcus aureus, and Enterococcus spp. A total of 145 surface samples were collected from six Italian hospitals and analyzed by both standard culture and LAMP methods, following two different incubation times (6 and 9 h) using pre-enrichment medium. Comparison with the reference method revealed that the LAMP assay achieved a sensitivity of 1.00 for all target pathogens at both 6 and 9 h of incubation. Specificity values were slightly higher at 6 h compared to 9 h: 0.93 vs 0.90 for P. aeruginosa, 0.91 vs 0.89 for Enterococcus spp., while remaining 0.92 for S. aureus, at both incubation times. These results suggest that a 6-h incubation period offers an optimal balance between speed and diagnostic accuracy, making LAMP a promising tool for rapid microbiological surveillance in healthcare settings.