Factors Impacting Persistence of Phi6 Bacteriophage, an Enveloped Virus Surrogate, on Fomite Surfaces

The persistence of Phi6 (Φ6) bacteriophage on surfaces commonly encountered in consumer-facing environments was evaluated. Φ6 has been utilized as a surrogate for enveloped viruses, including SARS-CoV-2—the causative agent of COVID-19—due to structural similarities, biosafety level 1 (BSL-1) status, and ease of use. Φ6 persistence on fomites was evaluated by characterizing the impact of the inoculum matrix (artificial saliva, phosphate-buffered saline [PBS], tripartite), inoculum level (low and high), and surface type (nonporous—aluminum, stainless steel, plastic, touchscreen, vinyl; porous—wood). Φ6 was inoculated onto surfaces at low and high inoculum levels for each inoculum matrix and incubated (20.54 ± 0.48°C) for up to 168 h. Φ6 was eluted from the surface and quantified via the double agar overlay assay to determine virus survival over time. For nonporous surfaces inoculated with artificial saliva and PBS, significantly higher D values were observed with high inoculum application according to the 95% confidence intervals. In artificial saliva, D values ranged from 1.00 to 1.35 h at a low inoculum and 4.44 to 7.05 h at a high inoculum across inoculation matrices and surfaces. D values for Φ6, regardless of the inoculum level, were significantly higher in tripartite than in artificial saliva and PBS for nonporous surfaces. In contrast with artificial saliva or PBS, D values in tripartite at low inoculum (D values ranging from 45.8 to 72.8 h) were greater than those at high inoculum (D values ranging from 26.4 to 45.5 h) on nonporous surfaces. This study characterized the impact of the inoculum matrix, inoculum level, and surface type on Φ6 survival on various surfaces relevant to fomite transmission in public settings. IMPORTANCE An important consideration in virus contact transmission is the transfer rate between hands and surfaces, which is driven by several factors, including virus persistence on inanimate surfaces. This research characterized Φ6 persistence on surfaces commonly encountered in public settings based on various factors. The inoculum matrix, which simulates the route of transmission, can impact virus persistence, and three separate matrices were evaluated in this study to determine the impact on Φ6 persistence over time. The number of microorganisms has also been suggested to impact persistence, which was evaluated here to simulate real-world contamination scenarios on six surface types. Results from this study will guide future research utilizing Φ6 or other surrogates for enveloped viruses of public health concern.