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

利用实时震荡诱导转化法对福尔马林固定石蜡包埋组织进行水提取并检测朊病毒病。

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Abstract

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.

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