Human variant Creutzfeldt-Jakob disease and sheep scrapie PrPres detection using seeded conversion of recombinant prion protein
Open Access
- 1 July 2009
- journal article
- Published by Oxford University Press (OUP) in Protein Engineering, Design and Selection
- Vol. 22 (8), 515-521
- https://doi.org/10.1093/protein/gzp031
Abstract
The pathological isoform of the prion protein (PrP(res)) can serve as a marker for prion diseases, but more practical tests are needed for preclinical diagnosis and sensitive detection of many prion infections. Previously we showed that the quaking-induced conversion (QuIC) assay can detect sub-femtogram levels of PrP(res) in scrapie-infected hamster brain tissue and distinguish cerebral spinal fluid (CSF) samples from normal and scrapie-infected hamsters. We now report the adaptation of the QuIC reaction to prion diseases of medical and agricultural interest: human variant Creutzfeldt-Jakob disease (vCJD) and sheep scrapie. PrP(res)-positive and -negative brain homogenates from humans and sheep were discriminated within 1-2 days with a sensitivity of 10-100 fg PrP(res). More importantly, in as little as 22 h we were able to distinguish CSF samples from scrapie-infected and uninfected sheep. These results suggest the presence of prions in CSF from scrapie-infected sheep. This new method enables the relatively rapid and sensitive detection of human CJD and sheep scrapie PrP(res) and may facilitate the development of practical preclinical diagnostic and high-throughput interference tests.Keywords
This publication has 29 references indexed in Scilit:
- Getting a Grip on Prions: Oligomers, Amyloids, and Pathological Membrane InteractionsAnnual Review of Biochemistry, 2009
- Human platelets as a substrate source for the in vitro amplification of the abnormal prion protein (PrPSc) associated with variant Creutzfeldt-Jakob diseaseTransfusion, 2009
- In vitro amplification of PrPSc derived from the brain and blood of sheep infected with scrapieJournal of General Virology, 2008
- Simplified ultrasensitive prion detection by recombinant PrP conversion with shakingNature Methods, 2008
- Mechanisms of prion protein assembly into amyloidProceedings of the National Academy of Sciences of the United States of America, 2008
- In vitro amplification and detection of variant Creutzfeldt–Jakob disease PrPScThe Journal of Pathology, 2007
- Ultrasensitive detection of scrapie prion protein using seeded conversion of recombinant prion proteinNature Methods, 2007
- Scrapie—Uncertainties, biology and molecular approachesBiochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 2007
- Formation of native prions from minimal components in vitroProceedings of the National Academy of Sciences, 2007
- How to find a prion: [URE3], [PSI+] and [β]Methods, 2006