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

IF 3.4 2区医学 Q1 CLINICAL NEUROLOGY

Neuropathology and Applied Neurobiology Pub Date : 2025-08-01 DOI:10.1111/nan.70028

Soňa Baranová, Radoslav Matěj, Jiri G Safar, Karel Holada

{"title":"Detection of Human Prion Seeding Activity in Formalin-Fixed Paraffin-Embedded Archival Tissues.","authors":"Soňa Baranová, Radoslav Matěj, Jiri G Safar, Karel Holada","doi":"10.1111/nan.70028","DOIUrl":null,"url":null,"abstract":"Aims: Formalin-fixed paraffin-embedded (FFPE) samples, routinely used in neuropathology, represent an invaluable resource for studying rare diseases like transmissible spongiform encephalopathies (TSE). Despite fixation-induced protein cross-linking, prion seeding activity can be effectively detected using the seeding amplification assays. In this study, we employed the second-generation real-time quaking-induced conversion (RT-QuIC) assay to analyse and quantify human prion seeding activity in FFPE brain tissues.Methods: FFPE frontal brain tissues were deparaffinised in xylene, followed by rehydration through descending concentrations of ethanol. The prion seeding activity in tissue homogenates was assessed by RT-QuIC assay utilising short recombinant hamster prion protein (rHaPrP90-231) as a substrate.Results: A total of 60 samples, including 30 cases of confirmed TSE, comprising both sporadic and genetic forms, as well as 30 non-TSE controls, were analysed. Prion seeding activity has been detected in all TSE samples except one sCJD (VV2) and one GSS (P102L) case, corresponding to an assay sensitivity of 93.3%. Conversely, we did not detect any RT-QuIC positivity in the control group, resulting in 100% specificity. The mean 50% prion seeding dose of FFPE sporadic TSE samples was 107.8/g of brain tissue.Conclusion: Our study emphasises high sensitivity and specificity of RT-QuIC assay for prion detection in archival human FFPE brain tissues and demonstrates its diagnostic reliability comparable to other tissue types even after years of storage. The applicability of FFPE samples in RT-QuIC assays facilitates retrospective diagnostics and provides logistical advantages for sample preservation and transportation.","PeriodicalId":19151,"journal":{"name":"Neuropathology and Applied Neurobiology","volume":"51 4","pages":"e70028"},"PeriodicalIF":3.4000,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12239053/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Neuropathology and Applied Neurobiology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1111/nan.70028","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CLINICAL NEUROLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

Aims: Formalin-fixed paraffin-embedded (FFPE) samples, routinely used in neuropathology, represent an invaluable resource for studying rare diseases like transmissible spongiform encephalopathies (TSE). Despite fixation-induced protein cross-linking, prion seeding activity can be effectively detected using the seeding amplification assays. In this study, we employed the second-generation real-time quaking-induced conversion (RT-QuIC) assay to analyse and quantify human prion seeding activity in FFPE brain tissues.

Methods: FFPE frontal brain tissues were deparaffinised in xylene, followed by rehydration through descending concentrations of ethanol. The prion seeding activity in tissue homogenates was assessed by RT-QuIC assay utilising short recombinant hamster prion protein (rHaPrP90-231) as a substrate.

Results: A total of 60 samples, including 30 cases of confirmed TSE, comprising both sporadic and genetic forms, as well as 30 non-TSE controls, were analysed. Prion seeding activity has been detected in all TSE samples except one sCJD (VV2) and one GSS (P102L) case, corresponding to an assay sensitivity of 93.3%. Conversely, we did not detect any RT-QuIC positivity in the control group, resulting in 100% specificity. The mean 50% prion seeding dose of FFPE sporadic TSE samples was 10^7.8/g of brain tissue.

Conclusion: Our study emphasises high sensitivity and specificity of RT-QuIC assay for prion detection in archival human FFPE brain tissues and demonstrates its diagnostic reliability comparable to other tissue types even after years of storage. The applicability of FFPE samples in RT-QuIC assays facilitates retrospective diagnostics and provides logistical advantages for sample preservation and transportation.

Abstract Image

查看原文本刊更多论文

人朊病毒在福尔马林固定石蜡包埋档案组织中的播种活性检测。

目的：福尔马林固定石蜡包埋（FFPE）样本通常用于神经病理学，是研究罕见疾病（如传染性海绵状脑病（TSE））的宝贵资源。尽管固定诱导的蛋白质交联，朊病毒的播种活性可以有效地检测使用播种扩增试验。在本研究中，我们采用第二代实时地震诱导转化（RT-QuIC）法分析和量化FFPE脑组织中人类朊病毒的播种活性。方法：将FFPE额叶脑组织在二甲苯中脱胶，然后通过降低浓度的乙醇补液。以重组短链仓鼠朊病毒蛋白（rHaPrP90-231）为底物，RT-QuIC法检测组织浆液中朊病毒的播种活性。结果：共分析了60份样本，其中包括30例确诊的TSE，包括散发型和遗传型，以及30例非TSE对照。除一例sCJD （VV2）和一例GSS （P102L）病例外，所有TSE样品均检测到朊病毒播种活性，检测灵敏度为93.3%。相反，我们在对照组中未检测到任何RT-QuIC阳性，因此特异性为100%。FFPE散发性TSE样本平均50%朊病毒播种剂量为107.8/g脑组织。结论：我们的研究强调了RT-QuIC法检测存档人类FFPE脑组织中朊病毒的高灵敏度和特异性，并且证明了其诊断可靠性，即使经过多年的储存也与其他组织类型相当。FFPE样品在RT-QuIC分析中的适用性促进了回顾性诊断，并为样品保存和运输提供了物流优势。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Neuropathology and Applied Neurobiology 医学-病理学

CiteScore

8.20

自引率

2.00%

发文量

审稿时长

6-12 weeks

期刊介绍： Neuropathology and Applied Neurobiology is an international journal for the publication of original papers, both clinical and experimental, on problems and pathological processes in neuropathology and muscle disease. Established in 1974, this reputable and well respected journal is an international journal sponsored by the British Neuropathological Society, one of the world leading societies for Neuropathology, pioneering research and scientific endeavour with a global membership base. Additionally members of the British Neuropathological Society get 50% off the cost of print colour on acceptance of their article.