title: 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: Laboratory of Persistent Viral Diseases, National Institute for Allergy and Infectious Diseases, National Institutes of Health, Rocky Mountain Laboratories, 903 S. 4th St., Hamilton, MT 59840, USA.
authors: Christina D
authors: CD
authors: Orrú
authors: Jason M
authors: JM
authors: Wilham
authors: Andrew G
authors: AG
authors: Hughson
authors: Lynne D
authors: LD
authors: Raymond
authors: Kristin L
authors: KL
authors: McNally
authors: Alex
authors: A
authors: Bossers
authors: Ciriaco
authors: C
authors: Ligios
authors: Byron
authors: B
authors: Caughey
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods*; D001923:Brain Chemistry; D007562:Creutzfeldt-Jakob Syndrome / Q000378:metabolism*; D006224:Cricetinae; D006801:Humans; D011328:Prions / Q000032:analysis* / Q000134:cerebrospinal fluid / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D012608:Scrapie / Q000378:metabolism*; D012756:Sheep
chemical_list: D011328:Prions; D011994:Recombinant Proteins