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

C.D. Orrú, J.M. Wilham, A.G. Hughson, L.D. Raymond, K.L. McNally, A. Bossers, C. Ligios, B. Caughey

Research output: Contribution to journalArticleAcademicpeer-review

50 Citations (Scopus)

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. 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. PrPres-positive and -negative brain homogenates from humans and sheep were discriminated within 1–2 days with a sensitivity of 10–100 fg PrPres. 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 PrPres and may facilitate the development of practical preclinical diagnostic and high-throughput interference tests.
Original languageEnglish
Pages (from-to)515-521
JournalProtein Engineering, Design & Selection
Volume22
Issue number8
DOIs
Publication statusPublished - 2009

Keywords

  • in-vitro amplification
  • misfolding cyclic amplification
  • cell-free formation
  • resistant state
  • ultrasensitive detection
  • cerebrospinal-fluid
  • prpsc
  • brain
  • form
  • binding

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