Use of rapid and innovative methods for environmental microbiological surveillance in hospital settings: real-time and on-site applications in the perspective of a mobile laboratory

Healthcare-associated infections (HAIs) remain a major challenge in hospital environments, where the presence of vulnerable patients and invasive procedures increases the risk of microbial transmission. Environmental microbiological surveillance plays a crucial role in identifying sources of contamination and supporting infection prevention strategies. However, conventional methods are often time-consuming and require centralized laboratory infrastructure, limiting their responsiveness and operational efficiency. This doctoral research aimed to develop and validate a rapid and reliable workflow for environmental microbiological surveillance in hospital settings. The study focused on two innovative analytical techniques: Loop-Mediated Isothermal Amplification (LAMP), for the direct and rapid detection of microbial DNA from environmental samples, and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS), for the fast identification of bacterial isolates. The LAMP study involved an initial optimization and adaptation of commercial kits originally designed for water testing, carried out under controlled laboratory conditions to evaluate their suitability for detecting pathogens on hospital surfaces, followed by subsequent field validation in hospital environments. Based on the promising results obtained in the first phases of the study, in collaboration with the manufacturer, new kits specifically tailored for hospital surface monitoring were developed and validated. In parallel, MALDI-TOF MS workflows were implemented based on the development of custom in-house spectral libraries to enhance identification accuracy in the context of hospital surveillance. The findings demonstrate the potential of these rapid methods to complement traditional approaches, reduce turnaround times, and improve the effectiveness of environmental monitoring programs. The integration of such technologies into mobile laboratory platforms is discussed as a future direction for decentralized hospital surveillance.