Comparison of acetone and sodium phosphotungstic acid precipitation for sample enrichment prior to RT-QuIC for the detection of prion disease

Authors: Nicholson, Eric; Veneziano, Susan

Submitted to: BMC Research Notes
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/25/2026
Publication Date: 3/19/2026
Citation: Nicholson, E.M., Veneziano, S.E. 2026. Comparison of acetone and sodium phosphotungstic acid precipitation for sample enrichment prior to RT-QuIC for the detection of prion disease. BMC Research Notes. https://doi.org/10.1186/s13104-026-07755-0.
DOI: https://doi.org/10.1186/s13104-026-07755-0

Interpretive Summary: The manuscript reports a method for enrichment of samples containing a low abundance of the disease associated form of the prion protein for detection of prion disease using an amplification based method to detect the disease. Prion diseases such as chronic wasting disease in deer and elk and scrapie in sheep and goats are fatal neurodegenerative diseases with a long incubation period and the potential to shed infectious particle in the environment long before the onset of clincally detectable disease. Detection has historically been limited to tissues that cannot be collected from live animals but recent advances in amplification based methods have afforded the ability to detect disease using fecal samples. The nature of these samples have, in some instances required enrichment using a multistep procedure. The method utilized here is a rapid single step procedure that meets and exceeds the current state of the art. This approach can be simply substituted in place of the existing enrichment procedure affording diagnostic labs a rapid and simple alternative for use in the early detection of prion disease from surveillance samples collected from wild and farmed animals.

Technical Abstract: Transmissible spongiform encephalopathies, also referred to as prion diseases, are fatal neurodegenerative diseases that are medically, economically, and ecologically important. In livestock and wildlife, detection to prevent introduction into a new herd or to assess environmental contamination is imperative for disease management. Real time quaking induced conversion (RT-QuIC) is an amplification-based approach with higher sensitivity than traditionally used immunoassays for TSEs and have been used to detected the presence of disease associated prions from various tissue, secretion, and excretion samples in both animals and humans. RT-QuIC also pairs well with upstream sample enrichment methods. In this study we compare the commonly used RT-QuIC sample enrichment, sodium phosphotungstic acid (NaPTA) precipitation, to acetone precipitation. Acetone precipitation is a widely used method for protein precipitation in the preparation and analysis of proteins but is less commonly used for prion disease sample analysis. This work shows that for brain samples, which typically contain a high amount of disease associated prion protein, acetone and NaPTA provide similar results to unenriched samples. In fecal samples, which have a low abundance of disease associated prion protein, acetone enrichment results in a higher degree of sensitivity than either NaPTA or a sample evaluated without enrichment. These data suggest that acetone enrichment can complement existing amplification based procedures affording diagnostic laboratories a rapid and simple sample enrichment approach in the early detection of prion disease from surveillance samples collected from wild and farmed animals.