Human variant Creutzfeldt-Jakob disease and sheep scrapie PrP(res) detection using seeded conversion of recombinant prion protein.

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.

• Authors: Christina D Orrú, Jason M Wilham, Andrew G Hughson, Lynne D Raymond, Kristin L McNally, Alex Bossers, Ciriaco Ligios, Byron Caughey
• Journal: Protein engineering, design & selection : PEDS (Protein Eng Des Sel) Vol. 22 Issue 8 Pg. 515-21 (Aug 2009) ISSN: 1741-0134 [Electronic] England
• PMID: 19570812 (Publication Type: Journal Article, Research Support, N.I.H., Intramural, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• Prions
• Recombinant Proteins

Topics

• Animals
• Biological Assay (methods)
• Brain Chemistry
• Creutzfeldt-Jakob Syndrome (metabolism)
• Cricetinae
• Humans
• Prions (analysis, cerebrospinal fluid, isolation & purification, metabolism)
• Recombinant Proteins (isolation & purification, metabolism)
• Scrapie (metabolism)
• Sheep