Estimating chronic wasting disease susceptibility in cervids using real-time quaking-induced conversion

Chronic wasting disease (CWD) is a prion-mediated, transmissible disease of cervids, including deer ( Odocoileus spp.), which is characterized by spongiform encephalopathy and death of the prion-infected animals. Official surveillance in the United States using immunohistochemistry (IHC) and ELISA entails the laborious collection of lymphoid and/or brainstem tissue after death. New, highly sensitive prion detection methods, such as real-time quaking-induced conversion (RT-QuIC), have shown promise in detecting abnormal prions from both antemortem and postmortem specimens. We compared RT-QuIC with ELISA and IHC for CWD detection utilizing deer retropharyngeal lymph node (RLN) tissues in a diagnostic laboratory setting. The RLNs were collected postmortem from hunter-harvested animals. RT-QuIC showed 100% sensitivity and specificity for 50 deer RLN (35 positive by both IHC and ELISA, 15 negative) included in our study. All deer were also genotyped for PRNP polymorphism. Most deer were homozygous at codons 95, 96, 116, and 226 (QQ/GG/AA/QQ genotype, with frequency 0.86), which are the codons implicated in disease susceptibility. Heterozygosity was noticed in Pennsylvania deer, albeit at a very low frequency, for codons 95GS (0.06) and 96QH (0.08), but deer with these genotypes were still found to be CWD prion-infected.

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Antemortem Detection of Chronic Wasting Disease Prions in Nasal Brush Collections and Rectal Biopsy Specimens from White-Tailed Deer by Real-Time Quaking-Induced Conversion

Journal of Clinical Microbiology ◽

10.1128/jcm.02699-15 ◽

2016 ◽

Vol 54 (4) ◽

pp. 1108-1116 ◽

Cited By ~ 27

Author(s):

Nicholas J. Haley ◽

Chris Siepker ◽

W. David Walter ◽

Bruce V. Thomsen ◽

Justin J. Greenlee ◽

...

Keyword(s):

Real Time ◽

Large Scale ◽

Chronic Wasting Disease ◽

Transmissible Spongiform Encephalopathy ◽

Rectal Biopsy ◽

Spongiform Encephalopathy ◽

Wasting Disease ◽

Experimental Conditions ◽

Biopsy Specimens

Chronic wasting disease (CWD), a transmissible spongiform encephalopathy of cervids, was first documented nearly 50 years ago in Colorado and Wyoming and has since spread to cervids in 23 states, two Canadian provinces, 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 of farmed or free-ranging deer and elk or surveillance studies of private or protected herds, where depopulation is contraindicated. This study sought to evaluate the sensitivity of the real-time quaking-induced conversion (RT-QuIC) assay by using recto-anal mucosa-associated lymphoid tissue (RAMALT) biopsy specimens and nasal brush samples collected antemortem from farmed white-tailed deer (n= 409). Antemortem findings were then compared to results from ante- and postmortem samples (RAMALT, brainstem, and medial retropharyngeal lymph nodes) evaluated by using the current gold standardin vitroassay, immunohistochemistry (IHC) analysis. We hypothesized that the sensitivity of RT-QuIC would be comparable to IHC analysis in antemortem tissues and would correlate with both the genotype and the stage of clinical disease. Our results showed that RAMALT testing by RT-QuIC assay had the highest sensitivity (69.8%) compared to that of postmortem testing, with a specificity of >93.9%. These data suggest that RT-QuIC, like IHC analysis, is an effective assay for detection of PrPCWDin rectal biopsy specimens and other antemortem samples and, with further research to identify more sensitive tissues, bodily fluids, or experimental conditions, has potential for large-scale and rapid automated testing for CWD diagnosis.

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Prion protein gene sequence and chronic wasting disease susceptibility in white-tailed deer (Odocoileus virginianus)

Prion ◽

10.1080/19336896.2015.1115179 ◽

2015 ◽

Vol 9 (6) ◽

pp. 449-462 ◽

Cited By ~ 15

Author(s):

Adam L Brandt ◽

Amy C Kelly ◽

Michelle L Green ◽

Paul Shelton ◽

Jan Novakofski ◽

...

Keyword(s):

Odocoileus Virginianus ◽

Prion Protein ◽

Gene Sequence ◽

Disease Susceptibility ◽

Protein Gene ◽

Chronic Wasting Disease ◽

Prion Protein Gene ◽

Wasting Disease

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Detection of Chronic Wasting Disease in the Lymph Nodes of Free-Ranging Cervids by Real-Time Quaking-Induced Conversion

Journal of Clinical Microbiology ◽

10.1128/jcm.01258-14 ◽

2014 ◽

Vol 52 (9) ◽

pp. 3237-3243 ◽

Cited By ~ 25

Author(s):

Nicholas J. Haley ◽

Scott Carver ◽

Laura L. Hoon-Hanks ◽

Davin M. Henderson ◽

Kristen A. Davenport ◽

...

Keyword(s):

Lymph Nodes ◽

Real Time ◽

Chronic Wasting Disease ◽

Wasting Disease ◽

Free Ranging

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Early detection of chronic wasting disease prions in urine of pre-symptomatic deer by real-time quaking-induced conversion assay

Prion ◽

10.4161/pri.24430 ◽

2013 ◽

Vol 7 (3) ◽

pp. 253-258 ◽

Cited By ~ 41

Author(s):

Theodore R. John ◽

Hermann M. Schätzl ◽

Sabine Gilch

Keyword(s):

Early Detection ◽

Real Time ◽

Chronic Wasting Disease ◽

Wasting Disease

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Evaluation of Winter Ticks (Dermacentor albipictus) Collected from North American Elk (Cervus canadensis) in an Area of Chronic Wasting Disease Endemicity for Evidence of PrP CWD Amplification Using Real-Time Quaking-Induced Conversion Assay

mSphere ◽

10.1128/msphere.00515-21 ◽

2021 ◽

Author(s):

N. J. Haley ◽

D. M. Henderson ◽

K. Senior ◽

M. Miller ◽

R. Donner

Keyword(s):

Real Time ◽

North American ◽

Chronic Wasting Disease ◽

Wasting Disease ◽

The Real ◽

First Finding ◽

North American Elk ◽

Dermacentor Albipictus ◽

Cervus Canadensis

This study reports the first finding of detectable levels of prions linked to chronic wasting disease in a tick collected from a clinically infected elk. Using the real-time quaking-induced conversion assay (RT-QuIC), “suspect” samples were also identified; these suspect ticks were more likely to have been collected from CWD-positive elk, though suspect amplification was also observed in ticks collected from CWD-negative elk.

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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

Journal of Clinical Microbiology ◽

10.1128/jcm.00947-18 ◽

2018 ◽

Vol 56 (9) ◽

Cited By ~ 9

Author(s):

Kristen A. Davenport ◽

Clare E. Hoover ◽

Nathaniel D. Denkers ◽

Candace K. Mathiason ◽

Edward A. Hoover

Keyword(s):

Real Time ◽

Protein Misfolding ◽

Chronic Wasting Disease ◽

Horizontal Transmission ◽

Wasting Disease ◽

Protein Misfolding Cyclic Amplification ◽

Rapid Spread ◽

Low Levels ◽

The Republic ◽

The Impact

ABSTRACT Chronic wasting disease (CWD), a fatal neurodegenerative prion disease of cervids, has spread across North America and has been detected in The Republic of Korea, Finland, and Norway. CWD appears to spread by horizontal transmission, and prions shed in saliva, feces, and urine are thought to contribute. However, studies investigating the rapid spread of CWD have been hampered by assay inhibitors and a lack of consistent and sensitive means to detect the relatively low levels of prions in these samples. Here we show that saliva frequently contains an inhibitor of the real-time quaking-induced conversion assay (RT-QuIC) and that the inhibitor is a member of the mucin family. To circumvent the inhibitor, we developed a modified protein misfolding cyclic amplification (PMCA) method to amplify CWD prions in saliva that were undetectable or ambiguous by RT-QuIC. Our results reinforce the impact of saliva in horizontal CWD transmission and highlight the importance of detection optimization.

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Detection of chronic wasting disease prion seeding activity in deer and elk feces by real-time quaking-induced conversion

Journal of General Virology ◽

10.1099/jgv.0.000844 ◽

2017 ◽

Vol 98 (7) ◽

pp. 1953-1962 ◽

Cited By ~ 11

Author(s):

Davin M. Henderson ◽

Joanne M. Tennant ◽

Nicholas J. Haley ◽

Nathaniel D. Denkers ◽

Candace K. Mathiason ◽

...

Keyword(s):

Real Time ◽

Chronic Wasting Disease ◽

Wasting Disease

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Real-Time Quaking-Induced Conversion Detection of PrPSc in Fecal Samples From Chronic Wasting Disease Infected White-Tailed Deer Using Bank Vole Substrate

Frontiers in Veterinary Science ◽

10.3389/fvets.2021.643754 ◽

2021 ◽

Vol 8 ◽

Author(s):

Soyoun Hwang ◽

Justin J. Greenlee ◽

Eric M. Nicholson

Keyword(s):

Real Time ◽

Chronic Wasting Disease ◽

Transmissible Spongiform Encephalopathy ◽

The United States ◽

Spongiform Encephalopathy ◽

Post Inoculation ◽

Sample Type ◽

Wasting Disease ◽

Fecal Samples ◽

Highly Sensitive Detection

Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy (TSE) that is fatal to free-range and captive cervids. CWD has been reported in the United States, Canada, South Korea, Norway, Finland, and Sweden, and the case numbers in both wild and farmed cervids are increasing rapidly. Studies indicate that lateral transmission of cervids likely occurs through the shedding of infectious prions in saliva, feces, urine, and blood into the environment. Therefore, the detection of CWD early in the incubation time is advantageous for disease management. In this study, we adapt real-time quacking-induced conversion (RT-QuIC) assays to detect the seeding activity of CWD prions in feces samples from clinical and preclinical white-tailed deer. By optimizing reaction conditions for temperature as well as the salt and salt concentration, prion seeding activity from both clinical and preclinical animals were detected by RT-QuIC. More specifically, all fecal samples collected from 6 to 30 months post inoculation showed seeding activity under the conditions of study. The combination of a highly sensitive detection tool paired with a sample type that may be collected non-invasively allows a useful tool to support CWD surveillance in wild and captive cervids.

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