Journal Article

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

Nicholson, Eric M., Hwang, Soyoun, Greenlee, Justin J.

2024

Overview

Objective The goal of the research presented here is to determine if methods previously developed for the aqueous extraction of PrP Sc from formalin-fixed paraffin-embedded tissue (FFPET) are applicable to the detection PrP Sc 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 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.

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Advancing Methods in Molecular Biology and Genetics / Animal diseases / Animals / Biochemical assays / Biomedical and Life Sciences / Biomedicine / Deer / Diagnosis / Diagnostic specimens / Extraction (Chemistry) / Formaldehyde / Formaldehyde - chemistry / Humans / Life Sciences / Medicine/Public Health / Methods / Mice / Paraffin / Paraffin Embedding - methods / Paraffin-embedded tissue / Prion / Prion diseases / Prion Diseases - diagnosis / Protection and preservation / Proteins / PrP / PrPSc Proteins - analysis / PrPSc Proteins - isolation & purification / PrPSc Proteins - metabolism / Research Note / RT-QuIC / Sheep / Spongiform encephalopathy / Tissue Fixation - methods / Tissues / Transmissible spongiform encephalopathy