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Detection of Human Prion Seeding Activity in Formalin-Fixed Paraffin-Embedded Archival Tissues

Soňa Baranová1, Radoslav Matěj2,3,4, Jiri G Safar5,6

  • 1Prion Research Laboratory, Institute of Medical Microbiology, First Faculty of Medicine, Charles University, Prague, Czech Republic.

Neuropathology and Applied Neurobiology
|July 9, 2025
PubMed
Abstract

Insights

The real-time quaking-induced conversion (RT-QuIC) assay effectively detects prion seeding activity in archival human formalin-fixed paraffin-embedded (FFPE) brain tissues. This method demonstrates high sensitivity and specificity for diagnosing transmissible spongiform encephalopathies (TSE).

Area of Science:

  • Neuropathology
  • Neurodegenerative Diseases
  • Biochemistry

Background:

  • Formalin-fixed paraffin-embedded (FFPE) tissues are crucial for studying rare diseases like transmissible spongiform encephalopathies (TSE).
  • Prion seeding activity detection in FFPE samples is challenging due to protein cross-linking but essential for diagnostics.

Purpose of the Study:

  • To employ the second-generation real-time quaking-induced conversion (RT-QuIC) assay for analyzing and quantifying human prion seeding activity in FFPE brain tissues.
  • To evaluate the diagnostic reliability of RT-QuIC for TSE detection in archival FFPE samples.

Main Methods:

  • FFPE brain tissues were deparaffinized and rehydrated.
  • Prion seeding activity was assessed using the RT-QuIC assay with recombinant hamster prion protein (rHaPrP90-231) substrate.

Main Results:

  • The RT-QuIC assay detected prion seeding activity in 93.3% of 30 confirmed TSE cases (sporadic and genetic forms).
  • The assay showed 100% specificity, with no positivity detected in 30 non-TSE control samples.
  • The mean 50% prion seeding dose in FFPE sporadic TSE samples was 10^7.8 per gram of brain tissue.

Conclusions:

  • RT-QuIC assay exhibits high sensitivity and specificity for prion detection in archival human FFPE brain tissues.
  • The assay's diagnostic reliability is comparable to other tissue types, even after prolonged storage.
  • Using FFPE samples in RT-QuIC assays offers advantages for retrospective diagnostics, sample preservation, and transportation.
Keywords:
FFPERT‐QuICSAAformalin‐fixed paraffin‐embeddedprionprion diseasereal‐time quaking‐induced conversion assayseeding amplification assay

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