Modified Protein Misfolding Cyclic Amplification Overcomes Real-Time Quaking-Induced Conversion Assay Inhibitors in Deer Saliva To Detect Chronic Wasting Disease Prions

Davenport, Kristen A.; Hoover, Clare; Denkers, Nathaniel D.; Mathiason, Candace K.; Hoover, Edward A.

doi:10.1128/jcm.00947-18

Cited by 32 publications

(37 citation statements)

References 52 publications

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“…Likewise, bioassays or cervid cell panel in vitro infection assays [35] on such RT-QuIC+/IHC-samples will also be valuable. Continued improvements in RT-QuIC methodology, enrichment protocols [36,37], and multi-laboratory confirmation assays on blinded sample panels will advance the use of amplification assays in CWD detection.…”

Section: Discussionmentioning

confidence: 99%

Progression of chronic wasting disease in white-tailed deer analyzed by serial biopsy RT-QuIC and immunohistochemistry

et al. 2020

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Chronic wasting disease (CWD) continues to spread or be recognized in the United States, Canada, and Europe. CWD is diagnosed by demonstration of the causative misfolded prion protein (PrP CWD) in either brain or lymphoid tissue using immunodetection methods, with immunohistochemistry (IHC) recognized as the gold standard. In recent years, in vitro amplification assays have been developed that can detect CWD prion seeding activity in tissues, excreta, and body fluids of affected cervids. These methods potentially offer earlier and more facile detection of CWD, both pre-and post-mortem. Here we provide a longitudinal profile of CWD infection progression, as assessed by both real-time quaking-induced conversion (RT-QuIC) and IHC on serial biopsies of mucosal lymphoid tissues of white-tailed deer orally exposed to low doses of CWD prions. We report that detection of CWD infection by RT-QuIC preceded that by IHC in both tonsil and recto-anal lymphoid tissue (RAMALT) in 14 of 19 deer (74%). Of the 322 biopsy samples collected in post-exposure longitudinal monitoring, positive RT-QuIC results were obtained for 146 samples, 91 of which (62%) were concurrently also IHC-positive. The lower frequency of IHC positivity was manifest most in the earlier post-exposure periods and in biopsies in which lymphoid follicles were not detected. For all deer in which RT-QuIC seeding activity was detected in a tonsil or RAMALT biopsy, PrP CWD was subsequently or concurrently detected by IHC. Overall, this study (a) provides a longitudinal profile of CWD infection in deer after low yet infectious oral prion exposure; (b) illustrates the value of RT-QuIC for sensitive detection of CWD; and (c) demonstrates an ultimate high degree of correlation between RT-QuIC and IHC positivity as CWD infection progresses.

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Section: Discussionmentioning

confidence: 99%

Progression of chronic wasting disease in white-tailed deer analyzed by serial biopsy RT-QuIC and immunohistochemistry

et al. 2020

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“…However, the complex biologic milieu in excreta contains assay inhibitors and/or non-specific activators that can interfere with in vitro amplification assays [26,27]. Nevertheless, through modifications of assay conditions, assays such as RT-QuIC can deliver sufficient sensitivity and specificity in problematic biologic samples [12,26,28,29].…”

Section: Introductionmentioning

confidence: 99%

Shedding and stability of CWD prion seeding activity in cervid feces

Tennant

Henderson

et al. 2020

PLoS ONE

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CWD is an emergent prion disease that now affects cervid species on three continents. CWD is efficiently spread in wild and captive populations, likely through both direct animal contact and environmental contamination. Here, by longitudinally assaying in feces of CWD-exposed white-tailed deer by RT-QuIC, we demonstrate fecal shedding of prion seeding activity months before onset of clinical symptoms and continuing throughout the disease course. We also examine the impact of simulated environmental conditions such as repeated freeze-thaw cycles and desiccation on fecal prion seeding activity. We found that while multiple (n = 7) freeze-thaw cycles substantially decreased fecal seeding activity, desiccation had little to no effect on seeding activity. Finally, we examined whether RT-QuIC testing of landscape fecal deposits could distinguish two premises with substantial known CWD prevalence from one in which no CWD-infected animals had been detected. In the above pilot study, this distinction was possible. We conclude that fecal shedding of CWD prions occurs over much of the disease course, that environmental factors influence prion seeding activity, and that it is feasible to detect fecal prion contamination using RT-QuIC.

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“…Each of the two methods has its advantages and disadvantages and may be sensitive to different inhibitors. For example, members of glycoprotein family mucins are potent inhibitors of RT-QuIC reaction [29]. This is a serious limitation for the analysis of saliva where mucins concentration is very high.…”

Section: Comparison Of Pmca With Real-time Quaking-induced Conversionmentioning

confidence: 99%

“…This is a serious limitation for the analysis of saliva where mucins concentration is very high. To circumvent the inhibition, Davenport and coauthors developed a modified PMCA with high detection sensitivity [29]. In another study endogenous polar lipids inhibited RT-QuIC reaction in brain samples.…”

Section: Comparison Of Pmca With Real-time Quaking-induced Conversionmentioning

confidence: 99%

Analysis of Protein Conformational Strains—A Key for New Diagnostic Methods of Human Diseases

Surguchov

2020

IJMS

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α-Synuclein is a naturally unfolded protein which easily aggregates and forms toxic inclusions and deposits. It is associated with several neurodegenerative diseases, including Parkinson’s disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA). These diseases, called synucleinopathies, have overlapping symptoms but require different methods of treatment. There are no reliable approaches for early diagnoses of these diseases, and as a result, the treatment begins late, and the disorders are often misdiagnosed. Recent studies revealed that α-synuclein forms distinctive spatial structures or strains at the early steps of these diseases, which may be used for early diagnosis. One of these early diagnostic methods called PMCA (protein misfolding cyclic amplification) allows identification of the distinct α-synuclein strains specific for different human diseases. The method is successfully used for differential diagnosis of patients with PD and MSA.

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Modified Protein Misfolding Cyclic Amplification Overcomes Real-Time Quaking-Induced Conversion Assay Inhibitors in Deer Saliva To Detect Chronic Wasting Disease Prions

Cited by 32 publications

References 52 publications

Progression of chronic wasting disease in white-tailed deer analyzed by serial biopsy RT-QuIC and immunohistochemistry

Progression of chronic wasting disease in white-tailed deer analyzed by serial biopsy RT-QuIC and immunohistochemistry

Shedding and stability of CWD prion seeding activity in cervid feces

Analysis of Protein Conformational Strains—A Key for New Diagnostic Methods of Human Diseases

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