1887

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Volume 96, Issue 1

Quantitative assessment of prion infectivity in tissues and body fluids by real-time quaking-induced conversion Free

Abstract

Prions are amyloid-forming proteins that cause transmissible spongiform encephalopathies through a process involving the templated conversion of the normal cellular prion protein (PrP) to a pathogenic misfolded conformation. Templated conversion has been modelled in several assays, including serial protein misfolding amplification, amyloid seeding and real-time quaking-induced conversion (RT-QuIC). As RT-QuIC measures formation of amyloid fibrils in real-time, it can be used to estimate the rate of seeded conversion. Here, we used samples from deer infected with chronic wasting disease (CWD) in RT-QuIC to show that serial dilution of prion seed was linearly related to the rate of amyloid formation over a range of 10 to 10 µg. We then used an amyloid formation rate standard curve derived from a bioassayed reference sample (CWD+ brain homogenate) to estimate the prion seed concentration and infectivity in tissues, body fluids and excreta. Using these methods, we estimated that urine and saliva from CWD-infected deer both contained 1–5 LD per 10 ml. Thus, over the 1–2 year course of an infection, a substantial environmental reservoir of CWD prion contamination accumulates.

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© Microbiology Society
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2015-01-01
2024-04-25
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Quantitative assessment of prion infectivity in tissues and body fluids by real-time quaking-induced conversion
J. Gen. Virol. 96, 210 (2015); https://doi.org/10.1099/vir.0.069906-0
/content/journal/jgv/10.1099/vir.0.069906-0
/content/journal/jgv/10.1099/vir.0.069906-0
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