Журнал микробных и биохимических технологий

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Application of Real-Time PCR to Identify Residual Bio-Decontamination of Confined Environments after Hydrogen Peroxide Vapor Treatment: Preliminary Results

Michele Pazienza, Maria Serena Britti, Mariachiara Carestia, Orlando Cenciarelli, Fabrizio D'Amico, Andrea Malizia, Carlo Bellecci, Pasquale Gaudio, Antonio Gucciardino, Mariarosa Bellino, Corrado Lancia, Annalaura Tamburrini and Roberto Fiorito

This study was conducted to assess the effectiveness of Hydrogen Peroxide Vapor (HPV) to remove biological contamination in a confined environment and to evaluate real-time PCR assay as a technique for the evaluation of the decontamination efficiency. Decontamination after the dispersion of biological aerosol is a main issue from a civilian, public health and military perspective. Despite the effectiveness of aggressive substances, eco-friendly but still efficient methods for decontamination are a relevant demand and Hydrogen Peroxide Vapor (HPV) is among the most recent and promising technologies in this field. Another related issue is: when an environment can be considered fully decontaminated? The answer clearly depends on the objectives of the decontamination and this will affect the choice of the methodology. Furthermore, classical microbiological and molecular biology techniques are commonly used to identify biological contamination and residual contamination, but many of them are time consuming and require advanced training for the operators who perform the analysis. This may represent a bottleneck, especially when a quick response to an emergency is needed (i.e. during an unconventional event like CBRNe ones). In this work, a combination of commercially available equipment for detection, identification and decontamination, was evaluated in partnership between the Italian Army, the Department of Industrial Engineering and the School of Medicine and Surgery of the University of Rome “Tor Vergata”. The purpose of this work was to find a setup for equipment and methodologies for detection, identification and decontamination, to implement in case of biological events. Preliminary results show that, despite the death of the microorganisms, nucleic acids are not completely degraded by HPV treatment and, as a consequence, that real-time PCR may be the adequate, quick and easy method to verify the efficiency of bio decontamination when nucleic acid degradation represent the final objective.