Method for evaluating the antiviral properties of materials under BSL-1 conditions

Although the global demand for antiviral materials is becoming ever more urgent, their development and screening is constrained by the requirement for high-biosafety facilities. We here propose a surrogate-based testing method capable of screening antiviral materials under Biosafety Level 1 (BSL-1) conditions using the enveloped bacteriophage phi 6 and the non-enveloped bacteriophage MS2 as surrogates for the highly pathogenic enveloped SARS-CoV-2 (BSL-3) and the non-enveloped enterovirus EV71 (BSL-2), respectively. While the relationship between the bacteriophage phi 6 and SARS-CoV-2 has been previously reported by our group, the work described here provides the first material-level validation of bacteriophage MS2 as a surrogate for enterovirus EV71 and combines both models in a unified broad-spectrum antiviral screening method. Using statistical correlation and regression analyses, validation across six representative materials previously characterized by our group revealed positive monotonic relationships and predictive trends between surrogate and pathogenic viruses at the material level. This method enables reproducible, quantitative and scalable screening of antiviral materials while accelerating discovery and substantially reducing cost, time, and biohazard waste. This article provides a detailed step-by-step protocol, including supplementary instructional videos, to facilitate its adoption and reproducibility in materials testing laboratories worldwide, regardless of their biosafety capabilities. • BSL-1 screening method that accelerates discovery of antiviral materials. • Broad-spectrum antiviral method (against enveloped and non-enveloped viruses). • Bacteriophages phi 6 and MS2 as surrogates of SARS-CoV-2 and enterovirus EV71. • Low-cost antiviral method validated across six different materials. • Step-by-step protocol supported by supplementary instructional videos.