PMCA to demonstrate prions inactivation method efficacy on reusable medical devices: a relevant alternative to animal bioassays

Abstract

Validation of prion inactivation processes for medical devices relies on in vivo experimental protocols. However, bioassays are costly, long (one to two years) and ethically disputable. Additionally, results obtained with one prion strain, for example 263K (hamster-adapted strain originating from sheep scrapie), cannot be easily extrapolated to relevant human prion strains, further questioning the utility of bioassays. Over the past two decades, cell-free prion amplification assays have emerged as potential alternatives to bioassays. Rather than measuring prion infectivity, they quantify prion seeding activity, i.e. the capacity to convert the normal prion protein into the disease-associated isoform. The results obtained by an optimized cell-free assay termed miniaturized-bead protein misfolding cyclic amplification (mb-PMCA), with four processes using three different prion strains, 263K and two human prions derived from variant and sporadic Creutzfeldt-Jakob diseases, were compared to published bioassays using the same three strains and processes, when available. Tests performed on reference processes (steam, sodium hydroxide, sodium hypochlorite) and low temperature H2O2 sterilization process (STERRAD NXTM Advanced cycle), showed perfect alignment between mb-PMCA and available bioassays. STERRAD NXTM Advanced cycle was efficacious on all three prion strains. These data confirm that PMCA and in particular mb-PMCA is a relevant alternative to animal bioassays for assessment of prion inactivation processes and the interest of some low temperature H2O2 sterilization cycles.