ABSTRACT
Aim: This study aimed to validate the efficacy of hydrogen peroxide vapor (HPV) decontamination technology set up in a biosafety level 3 (BSL-3) laboratory on surrogates and hazard group 3 (HG3) agents. Methods and Results: The HPV decontamination system (Bioquell) was assessed with both qualitative and quantitative methods on (1) spore surrogates (Geobacillus stearothermophilus, Bacillus atrophaeus, and Bacillus thuringiensis) in the BSL-3 laboratory and in the material airlock and on (2) HG3 agents (Bacillus anthracis; SARS-CoV-2, Venezuelan equine encephalitis virus [VEE], and Vaccinia virus) in the BSL-3 laboratory. Other HG3 bacteria likely to be handled in the BSL-3 laboratory (Yersinia pestis, Burkholderia mallei, Brucella melitensis, and Francisella tularensis) were excluded from the HPV decontamination assays as preliminary viability tests demonstrated the total inactivation of these agents after 48 h drying on different materials. Conclusions: The efficacy of HPV decontamination was validated with a reduction in viability of 5-7 log10 for the spores (surrogates and B. anthracis), and for the enveloped RNA viruses. Vaccinia showed a higher resistance to the decontamination process, being dependent on the biological indicator location in the BSL-3 laboratory.