{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,26]],"date-time":"2025-07-26T08:49:01Z","timestamp":1753519741544,"version":"3.37.3"},"reference-count":35,"publisher":"Public Library of Science (PLoS)","issue":"10","license":[{"start":{"date-parts":[[2024,10,23]],"date-time":"2024-10-23T00:00:00Z","timestamp":1729641600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["5R01NS061902-12"],"award-info":[{"award-number":["5R01NS061902-12"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["P01AI077774-11"],"award-info":[{"award-number":["P01AI077774-11"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["5R01AI093634-02"],"award-info":[{"award-number":["5R01AI093634-02"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["5R01AI156037-03"],"award-info":[{"award-number":["5R01AI156037-03"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["5R01NS107246-05"],"award-info":[{"award-number":["5R01NS107246-05"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["5R01AI112956-09"],"award-info":[{"award-number":["5R01AI112956-09"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Zoonotic transmission of bovine spongiform encephalopathy or mad cow disease, by presumed consumption of infected beef, has increased awareness of the public health risk associated with prion diseases. Chronic wasting disease (CWD) affects moose, elk, and deer, all of which are frequently consumed by humans. Clear evidence of CWD transmission to humans has not been demonstrated, yet, establishing whether CWD prions are present in muscle tissue preferentially consumed by humans is of increasing interest. Conventional assays including immunohistochemistry (IHC) and enzyme-linked immunosorbent assay (ELISA) lack the sensitivity to detect low concentrations of prions presumed to be present outside neural or lymphatic tissues. Here we combined two prion amplification assays, the product of protein misfolding cyclic amplification (PMCA) applied directly into real-time quaking induced conversion (RT-QuIC) [denoted now as PQ] to demonstrate the presence of prion seeding activity (i.e. prions) in ~55% of hamstring muscles harvested from CWD-positive white-tailed deer. This compares to prion detection in only 10% of the same samples employing standard RT-QuIC. To determine the extent of CWD dissemination within muscle tissues commonly consumed we tested 7 additional muscles from a subset of deer by PQ. Tongue demonstrated the highest level of prions with ~92% positive. All negative controls remained negative in all PMCA and RT-QuIC assays. We conclude that the combination of PMCA with RT-QuIC readout permits detection of low prion concentrations present in muscle tissue of CWD-infected deer. These findings further demonstrate the utility of amplification assays as tools to detect very low levels of prion burden and supports their use to fill knowledge gaps in our understanding of CWD pathogenesis and zoonotic potential.<\/jats:p>","DOI":"10.1371\/journal.pone.0309918","type":"journal-article","created":{"date-parts":[[2024,10,23]],"date-time":"2024-10-23T17:36:22Z","timestamp":1729704982000},"page":"e0309918","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":1,"title":["Enhanced detection of chronic wasting disease in muscle tissue harvested from infected white-tailed deer employing combined prion amplification assays"],"prefix":"10.1371","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1994-4947","authenticated-orcid":true,"given":"Caitlyn N.","family":"Kraft","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David W.","family":"Bissinger","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1918-6207","authenticated-orcid":true,"given":"Erin E.","family":"McNulty","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nathaniel D.","family":"Denkers","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Candace K.","family":"Mathiason","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2024,10,23]]},"reference":[{"issue":"3","key":"pone.0309918.ref001","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1016\/0022-510X(79)90171-0","article-title":"Familial Creutzfeldt-Jakob disease","volume":"42","author":"M Haltia","year":"1979","journal-title":"J Neurol Sci"},{"issue":"6","key":"pone.0309918.ref002","doi-asserted-by":"crossref","first-page":"901","DOI":"10.3201\/eid1806.120116","article-title":"Iatrogenic Creutzfeldt-Jakob disease, final assessment","volume":"18","author":"P Brown","year":"2012","journal-title":"Emerg Infect Dis"},{"issue":"11","key":"pone.0309918.ref003","doi-asserted-by":"crossref","DOI":"10.3390\/pathogens10111413","article-title":"Variant CJD: Reflections a Quarter of a Century on.","volume":"10","author":"DL Ritchie","year":"2021","journal-title":"Pathogens"},{"journal-title":"Expanding Distribution of Chronic Wasting Disease","author":"Center NWH.","key":"pone.0309918.ref004"},{"issue":"6","key":"pone.0309918.ref005","doi-asserted-by":"crossref","first-page":"e5916","DOI":"10.1371\/journal.pone.0005916","article-title":"Infectious prions in pre-clinical deer and transmission of chronic wasting disease solely by environmental exposure.","volume":"4","author":"CK Mathiason","year":"2009","journal-title":"PloS one"},{"issue":"3","key":"pone.0309918.ref006","doi-asserted-by":"crossref","first-page":"e0227094","DOI":"10.1371\/journal.pone.0227094","article-title":"Shedding and stability of CWD prion seeding activity in cervid feces.","volume":"15","author":"JM Tennant","year":"2020","journal-title":"PloS one."},{"issue":"1","key":"pone.0309918.ref007","doi-asserted-by":"crossref","DOI":"10.1099\/jgv.0.001825","article-title":"Longitudinal detection of prion shedding in nasal secretions of CWD-infected white-tailed deer","volume":"104","author":"CN Kraft","year":"2023","journal-title":"J Gen Virol"},{"key":"pone.0309918.ref008","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1093\/bmb\/66.1.185","article-title":"Bovine spongiform encephalopathy (BSE) and its epidemiology.","volume":"66","author":"PG Smith","year":"2003","journal-title":"Br Med Bull"},{"issue":"2","key":"pone.0309918.ref009","doi-asserted-by":"crossref","first-page":"e31449","DOI":"10.1371\/journal.pone.0031449","article-title":"Infectivity in skeletal muscle of cattle with atypical bovine spongiform encephalopathy.","volume":"7","author":"S Suardi","year":"2012","journal-title":"PloS one."},{"key":"pone.0309918.ref010","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1186\/1746-6148-8-22","article-title":"Experimental H-type and L-type bovine spongiform encephalopathy in cattle: observation of two clinical syndromes and diagnostic challenges","volume":"8","author":"T Konold","year":"2012","journal-title":"BMC Vet Res"},{"issue":"1","key":"pone.0309918.ref011","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1292\/jvms.13-0363","article-title":"The presence of disease-associated prion protein in skeletal muscle of cattle infected with classical bovine spongiform encephalopathy","volume":"76","author":"H Okada","year":"2014","journal-title":"J Vet Med Sci"},{"issue":"Pt 11","key":"pone.0309918.ref012","doi-asserted-by":"crossref","first-page":"3443","DOI":"10.1099\/vir.0.81777-0","article-title":"Prion protein in cardiac muscle of elk (Cervus elaphus nelsoni) and white-tailed deer (Odocoileus virginianus) infected with chronic wasting disease.","volume":"87","author":"JE Jewell","year":"2006","journal-title":"J Gen Virol"},{"issue":"4","key":"pone.0309918.ref013","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1177\/03009858231173467","article-title":"Distribution of the misfolded isoform of the prion protein in peripheral tissues and spinal cord of Rocky Mountain elk (Cervus elaphus nelsoni) with naturally occurring chronic wasting disease.","volume":"60","author":"TR Spraker","year":"2023","journal-title":"Vet Pathol"},{"issue":"5764","key":"pone.0309918.ref014","doi-asserted-by":"crossref","first-page":"1117","DOI":"10.1126\/science.1122864","article-title":"Prions in skeletal muscles of deer with chronic 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Review.","volume":"12","author":"MA Tranulis","year":"2023","journal-title":"Foods"},{"issue":"5","key":"pone.0309918.ref031","doi-asserted-by":"crossref","first-page":"e0216621","DOI":"10.1371\/journal.pone.0216621","article-title":"Comparison of conventional, amplification and bio-assay detection methods for a chronic wasting disease inoculum pool.","volume":"14","author":"E McNulty","year":"2019","journal-title":"PloS one."},{"key":"pone.0309918.ref032","doi-asserted-by":"crossref","first-page":"25098","DOI":"10.1038\/srep25098","article-title":"Detection and Quantification of CWD Prions in Fixed Paraffin Embedded Tissues by Real-Time Quaking-Induced Conversion.","volume":"6","author":"CE Hoover","year":"2016","journal-title":"Sci Rep."},{"issue":"1","key":"pone.0309918.ref033","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1128\/JVI.77.1.583-591.2003","article-title":"Rapid prion neuroinvasion following tongue infection","volume":"77","author":"JC Bartz","year":"2003","journal-title":"Journal of virology"},{"issue":"13","key":"pone.0309918.ref034","doi-asserted-by":"crossref","first-page":"6792","DOI":"10.1128\/JVI.78.13.6792-6798.2004","article-title":"Prion infection of skeletal muscle cells and papillae in the tongue","volume":"78","author":"ER Mulcahy","year":"2004","journal-title":"Journal of virology"},{"key":"pone.0309918.ref035","article-title":"Temporal characterization of prion shedding in secreta during a longitudinal study of chronic wasting disease in whtie-tailed deer","author":"N Denkers","year":"2024","journal-title":"Emerg Infect Dis"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0309918","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,10,23]],"date-time":"2024-10-23T17:36:32Z","timestamp":1729704992000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0309918"}},"subtitle":[],"editor":[{"given":"Byron","family":"Caughey","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2024,10,23]]},"references-count":35,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2024,10,23]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0309918","relation":{},"ISSN":["1932-6203"],"issn-type":[{"type":"electronic","value":"1932-6203"}],"subject":[],"published":{"date-parts":[[2024,10,23]]}}}