The digital prion infectivity and concentration assay
2012: 1 year / Grant: 30 000 euros
Project: The aim of dPica was to establish a new method of prion infectivity evaluation based on Poisson law distribution. The method is a sort of scrapie cell assay adapted for high-throughput automated screening in 96 or 384 wells. To achieve this task, several steps were proposed: - establishing a homogenous-phase time-resolved fluorescence resonance energy transfer (TR-FRET) based immunoassay for prion detection adapted for automation, -establishing a miniaturized cellbased prion infection, and incubation, -performing repeated mesurements, -evaluating the titer based on the Poisson distribution model without serial dilutions.
Main results and related published data: Many conditions have been tested for each step of the project and the development of a fully automated dPICA is in progress.
Falsig J, Sonati T, Herrmann US, Saban D, Li B, Arroyo K, Ballmer B, Liberski PP, Aguzzi A. (2012 November) Prion pathogenesis is faithfully reproduced in cerebellar organotypic slice cultures. PLoS Pathog. 8: e1002985.