Enhanced prion detection in biological samples by magnetic particle extraction and real-time quaking-induced conversion.

ABSTRACT

Prions have been demonstrated in body fluids and excreta using bioassay, but at levels too low for detection by conventional direct-detection assays. More rapid and sensitive detection of prions in these clinically accessible specimens would be valuable for diagnosis and investigations of transmission, environmental impact, and interventions. In addition to very low concentrations of prions, in vitro amplification assays are challenged by the presence of inhibitors in these complex sources. Here, we leverage the prion attribute of avid metal binding with the versatile power of real-time quaking-induced conversion (RT-QuIC) to enhance and simplify detection of chronic wasting-disease prions in biological samples. Iron oxide particle binding and magnetic extraction combined with RT-QuIC permitted rapid analysis of the low concentrations of prions in saliva, urine, faeces, and cerebrospinal fluid. These methods are pertinent to ante-mortem detection, monitoring, and surveillance, and could conceivably be applicable to other protein-misfolding disorders.