Real-time quaking induced conversion assay for prion seeding
Prions are pathogens that transmit their abnormal folding to normal prion protein. They induce fatal neurodegenerative diseases such as Creutzfeldt-Jakob disease. The real-time quaking induced conversion (RT-QuIC) assay helps to study prions and is an alternative to cost and time intensive animal studies.
The assay uses recombinant prion protein to which potentially infectious tissue homogenate is added. If the tissue has prion seeding activity, it induces aggregation in recombinant protein which can be monitored by the fluorophore Thioflavin T (ThT). For aggregation to occur, intermittent double-orbital shaking at 42 °C is required over an assay duration of up to 68 h. ThT fluorescence is acquired every 15 min to report on aggregation status.
This note proves the robustness of BMG LABTECH's Omega series required to perform the demanding assay. The microplate reader easily determined the 50 % seeding dose (SD50) of hamster scrapie brain homogenates.