Protocol for mpox virus inactivation in containment level 3 for safe handling of animal samples.

High biocontainment is required to handle infectious mpox virus (MPXV) specimens, which can hinder research and diagnostic efforts during outbreaks. Here, we present a protocol for inactivating MPXV-contaminated animal samples using commercial lysis buffers. We describe steps for virus propagation and quantification and MPXV inactivation. We then detail procedures for validating the inactivation using a cell-based technique and qPCR. This protocol provides a framework for safely managing MPXV in lower-containment laboratories, facilitating downstream applications and improved outbreak management.