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Prion detection by an amyloid seeding assay
Journal article   Open access   Peer reviewed

Prion detection by an amyloid seeding assay

David W Colby, Qiang Zhang, Shuyi Wang, Darlene Groth, Giuseppe Legname, Detlev Riesner and Stanley B Prusiner
Proceedings of the National Academy of Sciences - PNAS, Vol.104(52), pp.20914-20919
12/26/2007
DOI: 10.1073/pnas.0710152105
PMCID: PMC2409241
PMID: 18096717
url
https://doi.org/10.1073/pnas.0710152105View
Published (Version of record) Open Access

Abstract

Polymerization of recombinant prion protein (recPrP), which was produced in bacteria, into amyloid fibers was accompanied by the acquisition of prion infectivity. We report here that partially purified preparations of prions seed the polymerization of recPrP into amyloid as detected by a fluorescence shift in the dye Thioflavin T. Our amyloid seeding assay (ASA) detected PrP Sc , the sole component of the prion, in brain samples from humans with sporadic Creutzfeldt–Jakob disease, as well as in rodents with experimental prion disease. The ASA detected a variety of prion strains passaged in both mice and hamsters. The sensitivity of the ASA varied with strain type; for hamster Sc237 prions, the limit of detection was ≈1 fg. Some prion strains consist largely of protease-sensitive PrP Sc (sPrP Sc ), and these strains were readily detected by ASA. Our studies show that the ASA provides an alternative methodology for detecting both sPrP Sc and protease-resistant PrP Sc that does not rely on protease digestion or immunodetection.
 Biological Sciences femtogram protease-sensitive PrPSc Thioflavin T prion protein

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