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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Virus and Prion Research » Research » Publications at this Location » Publication #398573
Research Project: Elucidating the Pathobiology and Transmission of Transmissible Spongiform Encephalopathies

Location: Virus and Prion Research

Title: Aqueous extraction of formalin-fixed paraffin-embedded tissue and detection of prion disease using real-time quaking-induced conversion

Author
item Nicholson, Eric
item Greenlee, Justin
item HWANG, SOYOUN - Former ARS Employee

Submitted to: BMC Research Notes
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/2/2024
Publication Date: 9/16/2024
Citation: Nicholson, E.M., Greenlee, J.J., Hwang, S. 2024. Aqueous extraction of formalin-fixed paraffin-embedded tissue and detection of prion disease using real-time quaking-induced conversion. BMC Research Notes. https://doi.org/10.1186/s13104-024-06886-6.
DOI: https://doi.org/10.1186/s13104-024-06886-6

Interpretive Summary: The research presented in this manuscript is an extension of previously published methods for the separation of biological samples for study using real-time quaking induced conversion (RT-QuIC) from paraffin in which the tissues are embedded for use in histology. The method is robust and the ability to detect prion disease in these samples agrees with the known prion disease status. Two limitations are noted, the first is also technically a strength in the sense that RT-QuIC is sufficiently sensitive that it can detect contamination on the blade used to cut the tissue section or the instruments used to manipulate them. The second limitation is that the sensitivity appears to be lower, but it could simply reflect a lower amount of actual tissue in the detection system. Overall, this method offers a means to extract a sample from paraffin sections that does not require and organic solvent that would result in a mixed chemical-biologic hazard.

Technical Abstract: Objective: The goal of the research presented here is to determine if methods previously developed for the aqueous extraction of PrPSc from formalin-fixed paraffin-embedded tissue (FFPET) are applicable to the detection PrPSc by real-time quaking induced conversion (RT-QuIC). Previous work has utilized aqueous extraction of FFPET for detection of transmissible spongiform encephalopathies (TSEs) utilizing western blot and ELISA. This research extends the range of suitable methods for detection of TSEs in FFPET to RT-QuIC, which is arguably the most sensitive method to detect TSEs. Results: We found complete agreement between the TSE status and the results from RT-QuIC of seeded with the aqueous extract of FFPET samples. The method affords the diagnostic assessment TSE status by RT-QuIC of FFPET without the use of organic solvents that would otherwise create a mixed chemical-biological waste for disposal.