Stability and Reproducibility Underscore Utility of RT-QuIC for Diagnosis of Creutzfeldt-Jakob Disease

Cramm, Maria; Schmitz, Matthias; Karch, André; Mitrová, E; Kühn, Franziska; Schroeder, Bjoern O.; Raeber, Alex J.; Varges, Daniela; Kim, Yong‐Sun; Satoh, Katsuya; Collins, Steven J.; Zerr, Inga

doi:10.1007/s12035-015-9133-2

Cited by 173 publications

(171 citation statements)

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“…This is highlighted by the significant progress made in the field of RT-QuIC analysis within the last several years, demonstrating its utility for the identification of prion seeding activity in a multitude of tissues, including CSF, urine, saliva, blood, brain, lymph node tissue, and nasal lavage fluid/swabs (21,38,39,47,(49)(50)(51)(52)(53)(54). Additionally, intraassay variability has proven to be low (23,61), and with the ability to run a large number of samples simultaneously, generating rapid (Ͻ24 h), quantitative results, RT-QuIC is a fast, easy, and user-friendly assay with potential for widespread application in CWD research and monitoring.…”

Section: Discussionmentioning

confidence: 99%

Seeded Amplification of Chronic Wasting Disease Prions in Nasal Brushings and Recto-anal Mucosa-Associated Lymphoid Tissues from Elk by Real-Time Quaking-Induced Conversion

et al. 2016

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f Chronic wasting disease (CWD), a transmissible spongiform encephalopathy of cervids, was first documented nearly 50 years ago in Colorado and Wyoming and has since been detected across North America and the Republic of Korea. The expansion of this disease makes the development of sensitive diagnostic assays and antemortem sampling techniques crucial for the mitigation of its spread; this is especially true in cases of relocation/reintroduction or prevalence studies of large or protected herds, where depopulation may be contraindicated. This study evaluated the sensitivity of the real-time quaking-induced conversion (RT-QuIC) assay of recto-anal mucosa-associated lymphoid tissue (RAMALT) biopsy specimens and nasal brushings collected antemortem. These findings were compared to results of immunohistochemistry (IHC) analysis of ante-and postmortem samples. RAMALT samples were collected from populations of farmed and free-ranging Rocky Mountain elk (Cervus elaphus nelsoni; n ‫؍‬ 323), and nasal brush samples were collected from a subpopulation of these animals (n ‫؍‬ 205). We hypothesized that the sensitivity of RT-QuIC would be comparable to that of IHC analysis of RAMALT and would correspond to that of IHC analysis of postmortem tissues. We found RAMALT sensitivity (77.3%) to be highly correlative between RT-QuIC and IHC analysis. Sensitivity was lower when testing nasal brushings (34%), though both RAMALT and nasal brush test sensitivities were dependent on both the PRNP genotype and disease progression determined by the obex score. These data suggest that RT-QuIC, like IHC analysis, is a relatively sensitive assay for detection of CWD prions in RAMALT biopsy specimens and, with further investigation, has potential for large-scale and rapid automated testing of antemortem samples for CWD.T ransmissible spongiform encephalopathies are a group of progressively fatal neurodegenerative diseases caused by infectious proteins known as prions (1). The pathogenesis of prion diseases involves conversion of the endogenous cellular prion protein (PrP C ) present within specific tissues to the abnormal, protease-resistant form (PrP res ) following exposure to an infectious dose of PrP res (1). Chronic wasting disease (CWD), a naturally occurring prion disease of white-tailed deer (Odocoileus virginianus), mule deer (Odocoileus hemionus), Rocky Mountain elk (Cervus elaphus nelsoni), and moose (Alces alces), is the only known prion disease affecting free-ranging, nondomestic animals (2, 3). CWD was first described nearly 50 years ago as a fatal, wasting, spongiform encephalopathy of cervids in Colorado and Wyoming (4). The disease has since been documented in 23 U.S. states, 2 Canadian provinces, and, via exportation of farmed cervids, the Republic of Korea (5-8). Four of the 23 states (Texas, Iowa, Pennsylvania, and Ohio) were considered CWD free prior to 2012, with primary cases in three of these states reportedly arising in farmed cervids (9-11). With the movement of cervids across state and national borders, these new ep...

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Section: Discussionmentioning

confidence: 99%

Seeded Amplification of Chronic Wasting Disease Prions in Nasal Brushings and Recto-anal Mucosa-Associated Lymphoid Tissues from Elk by Real-Time Quaking-Induced Conversion

et al. 2016

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“…Following the report by Atarashi and colleagues 14) other groups have reported similar success with the RT-QuIC assay as a pre-mortem diagnostic test using CSF specimens [15][16][17] . In the study by McGuire and colleagues 16) , a blinded retrospective analysis of CSF samples from 108 patients yielded 91% sensitivity and 98% specificity, while their prospective study of 118 patients delivered 87% sensitivity and 100% specificity, with rec-huPrP evincing greater tendency to spontaneous conversion to PrP D in non-CJD cases than rec-haPrP.…”

Section: The Rt-quic As the "Next Generation" Pre-mortem Csf Diagnostmentioning

confidence: 73%

“…Using 12 CJD and 6 control CSF samples, this study additionally assessed the potential influence of storage temperatures (room temperature and +4 °C for up to 8 days; −80 °C for up to 9 years) and repeated freeze-thawing on the RT-QuIC assay with no significant adverse effects. In contrast, this study found that lysis of spiked red blood cells (RBCs) could adversely affect the sensitivity of the RT-QuIC either through spontaneous lysis over time (5,000 RBCs /µl >3 days) or if >1250 RBCs/µl were immediately sonicated 15) . In the most recent, larger collaborative study, 11 laboratories from 8 countries participated in a "blinded" ring trial of 15 CSF samples; there was perfect concordance between the laboratories for all CSF samples (McGuire et al Ann Neurol − in press).…”

Section: International Collaborative Studies To Validate the Csf Rt-qmentioning

confidence: 87%

“…In the study by McGuire and colleagues 16) , a blinded retrospective analysis of CSF samples from 108 patients yielded 91% sensitivity and 98% specificity, while their prospective study of 118 patients delivered 87% sensitivity and 100% specificity, with rec-huPrP evincing greater tendency to spontaneous conversion to PrP D in non-CJD cases than rec-haPrP. In the study of Cramm and colleagues 15) , CSF from 110 prion disease patients (including 39 genetic CJD) and 400 non-CJD controls (including 200 prospective samples) were analysed, delivering an overall 85% sensitivity and 99% specificity. Finally, in the report from Orrú and co-workers 17) , they describe significant refinements to the RT-QuIC assay, including use of truncated recombinant hamster PrP 90-231 (rec-thaPrP), addition of 0.002% SDS to CSF samples and an increased temperature from 42 °C to 55 °C, overall providing considerable shortening of the assay to 4-14 hours to observe a positive result.…”

Section: The Rt-quic As the "Next Generation" Pre-mortem Csf Diagnostmentioning

confidence: 99%

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RT-QuIC Assays in Humans … and Animals.

Collins

Sarros

2016

Food Safety

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Prion diseases are neurodegenerative diseases affecting both humans and animal species. The phenotypic spectrum is broad and includes Creutzfeldt-Jakob disease (CJD) and its variant zoonotic form (vCJD) in humans, while in animals, scrapie of sheep and goats, bovine spongiform encephalopathy and chronic wasting disease of deer, elk and moose are naturally occurring forms. Transmission and pathogenesis appear causally linked to the misfolding of the normal form of the prion protein (PrP C ) into disease associated conformers (PrP D ), the latter enriched in β-strand secondary structure. Over the past 10 years two protein amplification techniques, the protein misfolding cyclic amplification (PMCA) assay and real-time quaking induced conversion (RT-QuIC) assay have been developed and successfully deployed in prion biology across a range of scientific and clinical applications, including generation of de novo prions, quantitation of prion infectivity and ultra-sensitive detection of PrP D . While PMCA utilises sonication to facilitate protein amplification, RTQuIC employs vigorous shaking to achieve this outcome, with both techniques sharing the ability to amplify miniscule quantities of PrP D seed present in various tissues and body fluids to levels detectable using routine biochemical methods. The enhanced specificity of the RT-QuIC for detection of PrP D in cerebrospinal fluid (CSF) has spawned international collaborations to rigorously assess and validate the assay for clinical diagnostic purposes. In parallel with collaborative CSF validation studies have been successful efforts to refine the RT-QuIC allowing its use for more accessible body fluids or tissues such as urine and nasal brushings, as well as promote higher sample throughput, shorten assay times and offer accurate quantification of PrP D even at levels below those detectable by animal bioassays. Animal studies support the generic capacity of the RT-QuIC for PrP D detection, underpinning the utility of this assay for studying prion disease and the high likelihood of inter-convertibility of technical refinements for human and animal use.

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“…The diagnostic sensitivity of RT-QuIC is between 82-96% and practically full specificity. Nonetheless, technique still not that improved to discriminate CJD subtypes in comparison to age matched healthy and non-demented controls [32][33][34][35][36]. However, recent reports also showed lower detection sensitivity in CJD subtypes interrelated to type 2 abnormal prion protein (PrPSc) (VV2, MV2 and MM2) than in typical MM1 subtype of CJD.…”

Section: Prpc and Prpscmentioning

confidence: 99%