Optimization of An Enveloped Virus Surrogate, Bacteriophage Phi6, Recovery from Hands

Surfaces contaminated with enveloped viruses, such as severe acute respiratory syndrome coronavirus 2 and influenza virus, can potentially spread illness via hand contact. Often, the efficacy of hand hygiene interventions relies on virus recovery from hands. However, the recovery of bacteriophage phi6 (Φ6), a recommended surrogate for enveloped viruses, from the entire hands using the ASTM E2011-21 standard has not been optimized. For Φ6 recovery from the hands, three eluents [lysogeny broth (LC), tryptic soy broth (TSB), and 1.5% beef extract (BE)] and three recovery methods [glove juice method (GJM), hand rinsing, and modified dish method] were examined. The effects of inoculum application on either the palmar surface or the whole hand were compared, and virus recovery was assessed under wet and dry conditions to identify the optimal combinations for maximizing Φ6 recovery. Statistical differences among methods, inoculum application, and recovery types were identified. While no statistical difference was observed among the eluents (P = 0.281), LC demonstrated the highest Φ6 recovery efficiency, while TSB and BE had comparable recoveries. Two-way interaction effects were observed between method type vs. application type (P ≤ 0.05), method type vs. recovery type (P ≤ 0.05), and application type vs. recovery type (P ≤ 0.05), indicating these factors influencing one another. Additionally, no Φ6 recovery was obtained for the dry basis recovery type and the GJM method type. Based on the present study, to maximize Φ6 recovery from the hands during hand hygiene studies, inoculum should be applied to the palmar surface and recovered while it is still wet using LC.