Antiviral Effects of Ozonated Water for Handwashing Against SARS-CoV-2 Omicron Variant Using the E1052-20 Test Method of the American Society of Testing Materials International

Abstract

ABSTRACT Alcohol-based hand rubs are generally recommended for infection control of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Hand hygiene agents, including ozonated water (OW), have been used as an alternative because alcohol-based products may cause skin damage. A few reports are available on the virucidal activity of OW against SARS-CoV-2; therefore, this study examined the inhibitory effect of 0.4, 1.0, and 4.0 ppm OW on the Omicron variant infection to VeroE6 cells expressing transmembrane protease serine 2 by calculating the median tissue culture infectious dose (TCID50) using the E1052–20 test method of the American Society of Testing Materials International. Additionally, viral RNA copies were measured in culture supernatants of control or OW. Each OW concentration demonstrated ≥5.6 log10 TCID50 reduction, corresponding to ≥99.999% reduction of infectious virus, while the control phosphate-buffered saline showed no inactivation effect. More than 5 log10 copies/mL reduction of viral RNA was observed in the culture supernatant of each OW compared with the control. These findings suggest that exposure of SARS-CoV-2 to at least 0.4 ppm OW for 5 s is sufficient for viral inactivation and that OW can be used as an alternative hand hygiene agent to prevent SARS-CoV-2 infection.