Abstract

The pathological isoform of the prion protein (PrPres) 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 PrPres in scrapie-infected hamster brain tissue and distinguish cerebral spinal fluid (CSF) samples from normal and scrapie-infected hamsters. During FY2009, we adapted the QuIC reaction to prion diseases of medical and agricultural interest: human variant Creutzfeldt-Jakob disease (vCJD) and sheep scrapie. Prion-positive and negative brain homogenate samples from humans and sheep were discriminated within 1-2 days with a sensitivity of 10-100 femtograms PrPres. More importantly, in as little as 32h we were able to distinguish CSF samples from scrapie-infected and uninfected sheep. This is the first time that PrPres is detected in scrapie positive derived CSF. This new method enables the relatively rapid and sensitive detection of human CJD and sheep scrapie PrPres and may facilitate the development of practical preclinical diagnostic tests.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIAAI001086-01
Application #
7964733
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
1
Fiscal Year
2009
Total Cost
$758,320
Indirect Cost

  Institution

Name
National Institute of Allergy and Infectious Diseases
Department
Type
DUNS #
City
State
Country
Zip Code

  Publications

Haley, Nicholas J; Richt, Jürgen A; Davenport, Kristen A et al. (2018) Design, implementation, and interpretation of amplification studies for prion detection. Prion 12:73-82
Groveman, Bradley R; Orrù, Christina D; Hughson, Andrew G et al. (2018) Rapid and ultra-sensitive quantitation of disease-associated ?-synuclein seeds in brain and cerebrospinal fluid by ?Syn RT-QuIC. Acta Neuropathol Commun 6:7
Orrú, Christina D; Yuan, Jue; Appleby, Brian S et al. (2017) Prion seeding activity and infectivity in skin samples from patients with sporadic Creutzfeldt-Jakob disease. Sci Transl Med 9:
Saijo, Eri; Ghetti, Bernardino; Zanusso, Gianluigi et al. (2017) Ultrasensitive and selective detection of 3-repeat tau seeding activity in Pick disease brain and cerebrospinal fluid. Acta Neuropathol 133:751-765
Foutz, Aaron; Appleby, Brian S; Hamlin, Clive et al. (2017) Diagnostic and prognostic value of human prion detection in cerebrospinal fluid. Ann Neurol 81:79-92
Orrù, Christina D; Groveman, Bradley R; Hughson, Andrew G et al. (2017) RT-QuIC Assays for Prion Disease Detection and Diagnostics. Methods Mol Biol 1658:185-203
Groveman, Bradley R; Orrú, Christina D; Hughson, Andrew G et al. (2017) Extended and direct evaluation of RT-QuIC assays for Creutzfeldt-Jakob disease diagnosis. Ann Clin Transl Neurol 4:139-144
Bongianni, Matilde; Orrù, Christina; Groveman, Bradley R et al. (2017) Diagnosis of Human Prion Disease Using Real-Time Quaking-Induced Conversion Testing of Olfactory Mucosa and Cerebrospinal Fluid Samples. JAMA Neurol 74:155-162
Schmitz, Matthias; Cramm, Maria; Llorens, Franc et al. (2016) The real-time quaking-induced conversion assay for detection of human prion disease and study of other protein misfolding diseases. Nat Protoc 11:2233-2242
Haley, Nicholas J; Siepker, Chris; Hoon-Hanks, Laura L et al. (2016) Seeded Amplification of Chronic Wasting Disease Prions in Nasal Brushings and Recto-anal Mucosa-Associated Lymphoid Tissues from Elk by Real-Time Quaking-Induced Conversion. J Clin Microbiol 54:1117-26

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