Sensitive detection of chronic wasting disease prions recovered from environmentally relevant surfaces

Yuan, Qi; Rowden, Gage; Wolf, Tamir; Schwabenlander, Marc; Larsen, Peter A.; Bartelt–Hunt, Shannon L.; Bartz, Jason C.

doi:10.1016/j.envint.2022.107347

Cited by 14 publications

(24 citation statements)

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“…However, a limitation of most existing assays is the inability to detect prions that are bound to materials that interfere with the assay or will not fit into an assay well. Two recent studies reported detection of CWD prions from flat surfaces by sampling with foam swabs that are sonicated to elute prion seeds that are then concentrated and analyzed by RT-QuIC and PMCA [ 19 , 40 ]. Here, we demonstrate a more direct approach for detecting a broad range of pathologic PrP, α-Syn and tau prions or prion-like aggregates on large surfaces by simply exposing a sampling media to a surface (e.g., a surgical instrument) and then transferring the solution into an RT-QuIC assay plate for analysis.…”

Section: Introductionmentioning

confidence: 99%

Sensitive detection of pathological seeds of α-synuclein, tau and prion protein on solid surfaces

Orrú,

Groveman,

Hughson

et al. 2024

PLoS Pathog

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Prions or prion-like aggregates such as those composed of PrP, α-synuclein, and tau are key features of proteinopathies such as prion, Parkinson’s and Alzheimer’s diseases, respectively. Their presence on solid surfaces may be biohazardous under some circumstances. PrP prions bound to solids are detectable by ultrasensitive real-time quaking-induced conversion (RT-QuIC) assays if the solids can be immersed in assay wells or transferred to pads. Here we show that PrP prions can remain detectable on steel wires for at least a year, or even after enzymatic cleaning and sterilization. We also show that contamination of larger objects with pathological seeds of α-synuclein, tau, and PrP can be detected by simply assaying a sampling medium that has been transiently applied to the surface. Human α-synuclein seeds in dementia with Lewy bodies brain tissue was detected by α-synuclein RT-QuIC after drying of tissue dilutions with concentrations as low as 10−6 onto stainless steel. Tau RT-QuIC detected tau seeding activity on steel exposed to Alzheimer’s disease brain tissue diluted as much as a billion fold. Prion RT-QuIC assays detected seeding activity on plates exposed to brain dilutions as extreme as 10−5–10−8 from prion-affected humans, sheep, cattle and cervids. Sampling medium collected from surgical instruments used in necropsies of sporadic Creutzfeldt-Jakob disease-infected transgenic mice was positive down to 10−6 dilution. Sensitivity for prion detection was not sacrificed by omitting the recombinant PrP substrate from the sampling medium during its application to a surface and subsequent storage as long as the substrate was added prior to performing the assay reaction. Our findings demonstrate practical prototypic surface RT-QuIC protocols for the highly sensitive detection of pathologic seeds of α-synuclein, tau, and PrP on solid objects.

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Section: Introductionmentioning

confidence: 99%

Sensitive detection of pathological seeds of α-synuclein, tau and prion protein on solid surfaces

Orrú,

Groveman,

Hughson

et al. 2024

PLoS Pathog

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“…Here, we describe a method for determining the CWD status of a given sample based on what we coined the maxpoint ratio (MPR) [ 58 ]. This method was adapted from Cheng et al [ 59 , 60 ] and corrects replicates for background fluorescence and normalizes data output between experiments.…”

Section: Introductionmentioning

confidence: 99%

Standardization of Data Analysis for RT-QuIC-Based Detection of Chronic Wasting Disease

Rowden

Picasso-Risso

et al. 2023

Pathogens

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Chronic wasting disease (CWD) is a disease affecting cervids and is caused by prions accumulating as pathogenic fibrils in lymphoid tissue and the central nervous system. Approaches for detecting CWD prions historically relied on antibody-based assays. However, recent advancements in protein amplification technology provided the foundation for a new class of CWD diagnostic tools. In particular, real-time quaking-induced conversion (RT-QuIC) has rapidly become a feasible option for CWD diagnosis. Despite its increased usage for CWD-focused research, there lacks a consensus regarding the interpretation of RT-QuIC data for diagnostic purposes. It is imperative then to identify a standardized and replicable method for determining CWD status from RT-QuIC data. Here, we assessed variables that could impact RT-QuIC results and explored the use of maxpoint ratios (maximumRFU/backgroundRFU) to improve the consistency of RT-QuIC analysis. We examined a variety of statistical analyses to retrospectively analyze CWD status based on RT-QuIC and ELISA results from 668 white-tailed deer lymph nodes. Our results revealed an MPR threshold of 2.0 for determining the rate of amyloid formation, and MPR analysis showed excellent agreement with independent ELISA results. These findings suggest that the use of MPR is a statistically viable option for normalizing between RT-QuIC experiments and defining CWD status.

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“…South Korea, Norway, and Finland have also reported CWD, but the reported prevalence in these locations is much lower [ 3 ]. At this point, eradication of CWD from endemic areas is extremely unlikely, as mis-folded prions such as CWD are very stable in the environment [ 4 , 5 ], and the infectious dose required for transmission to additional deer is extremely low [ 6 ]. As CWD continues to spread across North America and other countries, the number of humans consuming CWD-infected meat and cervid products will increase.…”

Section: Introductionmentioning

confidence: 99%

Second passage experiments of chronic wasting disease in transgenic mice overexpressing human prion protein

Race

Baune

Williams

et al. 2022

Vet Res

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Chronic wasting disease (CWD) is a prion disease of cervids including deer, elk, reindeer, and moose. Human consumption of cervids is common, therefore assessing the risk potential of CWD transmission to humans is critical. In a previous study, we tested CWD transmission via intracerebral inoculation into transgenic mice (tg66 and tgRM) that over-expressed human prion protein. Mice screened by traditional prion detection assays were negative. However, in a group of 88 mice screened by the ultrasensitive RT-QuIC assay, we identified 4 tg66 mice that produced inconsistent positive RT-QuIC reactions. These data could be false positive reactions, residual input inoculum or indicative of subclinical infections suggestive of cross species transmission of CWD to humans. Additional experiments were required to understand the nature of the prion seeding activity in this model. In this manuscript, second passage experiments using brains from mice with weak prion seeding activity showed they were not infectious to additional recipient tg66 mice. Clearance experiments showed that input CWD prion seeding activity was eliminated by 180 days in tg66 mice and PrPKO mice, which are unable to replicate prion protein, indicating that the weak positive levels of seeding activity detected at later time points was not likely residual inoculum. The failure of CWD prions to cause disease in tg66 after two sequential passages suggested that a strong species barrier prevented CWD infection of mice expressing human prion protein.

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Sensitive detection of chronic wasting disease prions recovered from environmentally relevant surfaces

Cited by 14 publications

References 50 publications

Sensitive detection of pathological seeds of α-synuclein, tau and prion protein on solid surfaces

Sensitive detection of pathological seeds of α-synuclein, tau and prion protein on solid surfaces

Standardization of Data Analysis for RT-QuIC-Based Detection of Chronic Wasting Disease

Second passage experiments of chronic wasting disease in transgenic mice overexpressing human prion protein

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