Chapter 19 – Prion protein amplification techniques

Protein amplification techniques exploit the ability of PrPTSE to induce a conformational change in prion protein (PrP) in a continuous fashion, so that the small amount of PrPTSE found in tissues and biologic fluids in prion diseases can be amplified to a point where they are detectable by conventional laboratory techniques. The most widely used protein aggregation assays are protein misfolding cyclic amplification assay (PMCA) and real-time quaking-induced conversion (RT-QuIC). These assays have been used extensively in both animal and human prion disease in studies ranging from the development of diagnostics, understanding disease transmission potential, to investigating mechanisms underlying neurodegeneration. In human prion disease, cerebrospinal fluid (CSF) RT-QuIC analysis has been shown to be a highly sensitive and specific test for sporadic Creutzfeldt-Jakob disease (sCJD) and has now been included in the diagnostic criteria. It is also a useful investigation for some genetic forms of prion disease where other cerebrospinal fluid tests may be negative. PMCA shows great potential for the diagnosis of variant CJD (vCJD) and has the ability to distinguish vCJD from sCJD, which may become increasingly important with emergence of a patient with neuropathologically confirmed vCJD associated with PRNP codon129MV, which indicates that a new wave of vCJD cases is likely and that these may be difficult to distinguish from sCJD.