I may be wrong but I didn't think they could test live animals.
CWD TESTING
Other diagnostic tests and technologies that allow more rapid testing of larger numbers of samples continue to be developed. A rectal associated mucosal lymphoid tissue (RAMALT) biopsy (live-animal) test has been developed by researchers and appears to hold promise for future use in certain CWD monitoring or management situations in farmed cervids. This technique utilizes the current IHC testing technologies described above.
*** The RAMALT is not currently approved as an official test for CWD.
https://www.aphis.usda.gov/aphis/ourfocus/animalhealth/animal-disease-information/sa_alternate_livestock/sa_cervid_health/sa_cwd/ct_diagnostics
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
Nicholas J. Haley#,a, Chris Siepkera, Laura L. Hoon-Hanksb, Gordon Mitchellc, W. David Walterd, Matteo Mancae, Ryan J. Monellof, Jenny G. Powersf, Margaret A. Wildf, Edward A. Hooverb, Byron Caugheye and Jürgen A. Richta + Author Affiliations
Department of Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University (KSU), Manhattan, KS, USAa Department of Microbiology, Immunology and Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, USAb Canadian Food Inspection Agency, National and OIE Reference Laboratory for Scrapie and CWD, Ottawa Laboratory Fallowfield, Ottawa, ON, Canadac U.S. Geological Survey, Pennsylvania Cooperative Fish and Wildlife Research Unit, The Pennsylvania State University, University Park, PA, USAd TSE/Prion Biochemistry Section, Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories (RML), National Institute of Allergy and Infectious Diseases, Hamilton, MT, USAe National Park Service, Biological Resources Division, 1201 Oak Ridge Drive, Suite 200, Fort Collins, Colorado 80525, USAf
ABSTRACT
Chronic wasting disease (CWD), a transmissible spongiform encephalopathy of cervids, was first documented nearly fifty years ago in Colorado and Wyoming, and has since been detected across North America and to 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 spread; this is especially true in cases of relocation/reintroduction, or prevalence studies in large or protected herds where depopulation may be contraindicated. This study sought to evaluate the sensitivity of the real-time quaking-induced conversion (RT-QuIC) assay in recto-anal mucosa associated lymphoid tissue (RAMALT) biopsies and nasal brushings collected antemortem. These findings were compared to results from ante- and postmortem samples evaluated using immunohistochemistry (IHC). RAMALT samples were collected from populations of farmed and free-ranging Rocky Mountain elk (Cervus elaphus nelsoni, n=323), with nasal brushes collected from a subpopulation of these animals (n=205). We hypothesized the sensitivity of RT-QuIC would be comparable to IHC in RAMALT, and would correspond to IHC of postmortem tissues. We found RAMALT sensitivity (77.3%) to be highly correlative between RT-QuIC and IHC. Sensitivity was lower when testing nasal brushings (34%), though both RAMALT and nasal brush sensitivities were dependent on both PRNP genotype and disease progression determined by obex score. These data suggest that RT-QuIC, like IHC, is a relatively sensitive assay for detection of CWD prions in RAMALT biopsies, and with further investigation has potential for large scale and rapid automated testing for CWD in antemortem samples.
FOOTNOTES
↵#Corresponding author (e-mail:
nicholas.j.haley@gmail.com) Copyright © 2016, American Society for Microbiology. All Rights Reserved.
http://jcm.asm.org/content/early/2016/02/11/JCM.02700-15.abstract
Saturday, February 20, 2016
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
http://chronic-wasting-disease.blogspot.com/2016/02/seeded-amplification-of-chronic-wasting.html
Sunday, February 14, 2016
Antemortem detection of chronic wasting disease prions in nasal brush collections and rectal biopsies from white-tailed deer by real time quaking-induced conversion
http://chronic-wasting-disease.blogspot.com/2016/02/antemortem-detection-of-chronic-wasting.html
Research Project: TRANSMISSION, DIFFERENTIATION, AND PATHOBIOLOGY OF TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHIES Title: Clinical stage of infection is critical in the antemortem diagnosis of chronic wasting disease in deer and elk
Authors
item Siepker, Chris - item Haley, Nicholas - item Walter, W. David - item Hoon-Hanks, Laura - item Monello, Ryan - item Powers, Jenny - item Greenlee, Justin item Thomsen, Bruce - item Lehmkuhl, Aaron - item Mitchell, Gordon - item Nichols, Tracy - item Hoover, Edward - item Richt, Juergen -
Submitted to: Prion Publication Type: Abstract Only Publication Acceptance Date: March 5, 2015 Publication Date: N/A
Technical Abstract: Chronic wasting disease (CWD) is an efficiently transmitted spongiform encephalopathy of cervids (e.g. deer, elk, and moose), and is the only known prion disease affecting both free-ranging wildlife and captive animals. The antemortem detection of CWD and other prion diseases has proven difficult, due in part to difficulties in identifying an appropriate peripheral tissue specimen and complications with conventional test sensitivity. At present, biopsies of the recto-anal mucosal-associated lymphoid tissues (RAMALT) have shown promising sensitivity and are practical to collect in live animals. Nasal brush collections have likewise proven both sensitive and practical for identification of prion infections in humans. In this study, we evaluated both RAMALT and nasal brush collections by real time quaking-induced conversion (RT-QuIC), and compared our findings to RAMALT immunohistochemistry as well as conventional postmortem evaluation of obex and retropharyngeal lymph node tissues from over 700 captive and free-ranging deer and elk in areas with endemic CWD. We correlated our results with various clinical findings, including pathological stage of infection as determined by obex scoring, PrP genotype, age, and sex. While the sensitivity of RAMALT RT-QuIC analyses exceeded that of RAMALT IHC (69-80% vs. >44%) and nasal brush collections (15-30%), the sensitivity of both biopsy and nasal brush analyses were dependent primarily on clinical stage of disease, although PrP genotype was also an important predictor of sample positivity. Our findings further demonstrate the potential and limitations of antemortem sample analyses by RT-QuIC in the identification and management of prion diseases.
http://www.ars.usda.gov/research/publications/publications.htm?SEQ_NO_115=314836
Research Project: TRANSMISSION, DIFFERENTIATION, AND PATHOBIOLOGY OF TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHIES
Title: Antemortem detection of chronic wasting disease prions in nasal brush collections and rectal biopsies from white-tailed deer by real time quaking-induced conversion
Authors
item Haley, Nicholas - item Siepker, Chris - item Walter, W. David - item Thomsen, Bruce - item Greenlee, Justin item Lehmkuhl, Aaron - item Richt, Jürgen -
Submitted to: Journal of Clinical Microbiology Publication Type: Peer Reviewed Journal Publication Acceptance Date: November 27, 2015 Publication Date: February 10, 2016 Citation: Haley, N.J., Siepker, C., Walter, W.D., Thomsen, B.V., Greenlee, J.J., Lehmkuhl, A.D., Richt, J.A. 2016. 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. 54(4):1108-1116.
Interpretive Summary: Chronic Wasting Disease (CWD), a fatal neurodegenerative disease that occurs in farmed and wild cervids (deer and elk) of North America, is a transmissible spongiform encephalopathy (TSE). TSEs are caused by infectious proteins called prions that are resistant to various methods of decontamination and environmental degradation. Early diagnosis of CWD in wild and captive herds would be very helpful to controlling the spread of CWD, for which there are not yet any preventative or treatment measures available. The purpose of this study was to test a laboratory method of prion detection (real-time Quaking Induced Conversion; RT-QuIC) that has the potential to detect very low levels of infectious prions in samples collected from live animals against the gold standard diagnostic where abnormal prion in tissues is stained on a microscope slide. This study reports that RT-QuIC detects more cases of CWD than standard methods, but also can identify a small number of animals without CWD as being positive. In the case of CWD, where it is likely that large numbers of animals within a herd may be positive, misidentifying a negative as a positive may have less of an impact than in the case of other prion diseases such as bovine spongiform encephalopathy considering that this test allows testing much larger numbers of samples with a faster turn around time than traditional methods. This information could have an impact on regulatory and wildlife officials developing plans to reduce or eliminate CWD and cervid farmers that want to ensure that their herd remains CWD-free. Technical Abstract: Chronic wasting disease (CWD), a transmissible spongiform encephalopathy of cervids, was first documented nearly fifty years ago in Colorado and Wyoming and has since spread to cervids in 23 states, 2 Canadian provinces, and the Republic of Korea. The increasing expansion of this disease makes the development of sensitive diagnostic assays and antemortem sampling techniques crucial for the mitigation of spread; this is especially true in cases of relocation/reintroduction of farmed or free-ranging deer and elk, or surveillance studies in private or protected herds where depopulation may be contraindicated. This study sought to evaluate the sensitivity of the real-time quaking-induced conversion (RT-QuIC) assay in samples collected antemortem. Antemortem findings were then compared to results from ante- and postmortem samples evaluated using the current gold standard diagnostic assay, immunohistochemistry (IHC). Recto-anal mucosal associated lymphoid tissue (RAMALT) biopsies and nasal brush collections from three separate herds of farmed white-tailed deer (n=409) were evaluated, along with standard postmortem microscopic analysis of brainstem at the level of the obex and retropharyngeal lymph nodes. We hypothesized the sensitivity of RT-QuIC would be comparable to IHC in antemortem tissues, and would correlate with both genotype and stage of clinical disease. Our results showed that RAMALT testing by RT-QuIC had the highest sensitivity (69.8%) when compared to postmortem testing. This data suggests that RT-QuIC, like IHC, is a fairly sensitive assay for detection of CWD prions in rectal biopsies and other antemortem samples, and with further investigation has potential for large scale and rapid automated testing for CWD diagnosis.
http://www.ars.usda.gov/research/publications/publications.htm?seq_no_115=320291
Research Project: TRANSMISSION, DIFFERENTIATION, AND PATHOBIOLOGY OF TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHIES
Title: Clinical stage of infection is critical in the antemortem diagnosis of chronic wasting disease in deer and elk
Authors
item Siepker, Chris - item Haley, Nicholas - item Walter, W. David - item Hoon-Hanks, Laura - item Monello, Ryan - item Powers, Jenny - item Greenlee, Justin item Thomsen, Bruce - item Lehmkuhl, Aaron - item Mitchell, Gordon - item Nichols, Tracy - item Hoover, Edward - item Richt, Juergen -
Submitted to: Prion Publication Type: Abstract Only Publication Acceptance Date: March 5, 2015 Publication Date: N/A
Technical Abstract: Chronic wasting disease (CWD) is an efficiently transmitted spongiform encephalopathy of cervids (e.g. deer, elk, and moose), and is the only known prion disease affecting both free-ranging wildlife and captive animals. The antemortem detection of CWD and other prion diseases has proven difficult, due in part to difficulties in identifying an appropriate peripheral tissue specimen and complications with conventional test sensitivity. At present, biopsies of the recto-anal mucosal-associated lymphoid tissues (RAMALT) have shown promising sensitivity and are practical to collect in live animals. Nasal brush collections have likewise proven both sensitive and practical for identification of prion infections in humans. In this study, we evaluated both RAMALT and nasal brush collections by real time quaking-induced conversion (RT-QuIC), and compared our findings to RAMALT immunohistochemistry as well as conventional postmortem evaluation of obex and retropharyngeal lymph node tissues from over 700 captive and free-ranging deer and elk in areas with endemic CWD. We correlated our results with various clinical findings, including pathological stage of infection as determined by obex scoring, PrP genotype, age, and sex. While the sensitivity of RAMALT RT-QuIC analyses exceeded that of RAMALT IHC (69-80% vs. >44%) and nasal brush collections (15-30%), the sensitivity of both biopsy and nasal brush analyses were dependent primarily on clinical stage of disease, although PrP genotype was also an important predictor of sample positivity. Our findings further demonstrate the potential and limitations of antemortem sample analyses by RT-QuIC in the identification and management of prion diseases.
http://www.ars.usda.gov/research/publications/publications.htm?SEQ_NO_115=314836
*** CHRONIC WASTING DISEASE TESTING PROTOCAL FOR WILD CERVIDAE proposing the United States Animal Health Association (USAHA) urge the USDA to amend CFR 81.3 ***
Committee Business:
One resolution was proposed by a committee member titled CHRONIC WASTING DISEASE TESTING PROTOCAL FOR WILD CERVIDAE proposing the United States Animal Health Association (USAHA) urge the USDA to amend CFR 81.3 (b); proposing wild cervids captured for interstate movement and release, have two forms of identification, one of which that is official identification, must be PrP genotyped for Chronic Wasting Disease resistance, tested for Chronic Wasting Disease using a rectal biopsy test. The committee discussed and debated the terms and science related to this resolution proposal including that currently there is no science indicating there are “genotype resistant” cervids to acquiring the CWD prion. The term “resistant” is miss-leading. There are only different cervid genotypes that acquire the infectious prions at different rates and show clinical signs at variable rates, some at prolonged periods after acquiring the prion or they are slow to accumulate detectable levels.. Since all infected animals would be presumed to be capable of shedding the prions into the environment, genotypes with clinical “resistance” or prolonged indication of clinical presentation of the disease, may well potentially be considered prolonged shedders of the prion. Additionally there was discussion put forth by several committee members concerning the lack of regulatory validation of the rectal biopsy test. Also, the test can only be used on young animals and there I significant test sensitivity and specificity variability between cervid species when using this test. A new motion to the proposed resolution was to table this resolution, reword the resolution potentially to be a recommendation for USDA to provide a guidance document to the states for surveillance of CWD on interstate translocations od wild cervids. It was proposed that this new resolution/recommendation be discussed during the Farmed Cervid Subcommittee and forward then to the Captive Wildlfie and Alternative Livestock committee. The motion was proposed by member Charlie Seale and seconded by member Sean Shaffer which was passed by committee. The Committee on Wildlife Diseases adjourned at 515 PM.
http://www.usaha.org/Portals/6/Reports/2015/report-wd-2015.pdf
Tuesday, September 22, 2015
*** Host Determinants of Prion Strain Diversity Independent of Prion Protein Genotype
http://transmissiblespongiformencephalopathy.blogspot.com/2015/09/host-determinants-of-prion-strain.html
Friday, August 28, 2015
*** Chronic Wasting Disease CWD TSE Prion Diagnostics and subclinical infection
http://chronic-wasting-disease.blogspot.com/2015/08/chronic-wasting-disease-cwd-tse-prion.html
Monday, July 18, 2016
Texas Parks Wildlife Dept TPWD HIDING TSE (CWD) in Deer Herds, Farmers Sampling Own Herds, Rapid Testing, False Negatives, a Recipe for Disaster
http://chronic-wasting-disease.blogspot.com/2016/07/texas-parks-wildlife-dept-tpwd-hiding.html
IOWA CHRONIC WASTING DISEASE CWD TSE PRION TOTAL TO DATE 304 CASES WILD AND CAPTIVE REPORT UPDATE JULY 2016
[SIZE=+0][/SIZE]
WILD 6 CWD POSITIVE TO DATE
CAPTIVE 298 CWD POSITIVE TO DATE
personal communications...tss
http://chronic-wasting-disease.blogspot.com/2016/07/iowa-chronic-wasting-disease-cwd-tse.html
Friday, February 05, 2016
*** Report of the Committee on Wildlife Diseases FY2015 CWD TSE PRION Detections in Farmed Cervids and Wild
http://chronic-wasting-disease.blogspot.com/2016/02/report-of-committee-on-wildlife.html
Sunday, July 17, 2016
CHRONIC WASTING DISEASE CWD TSE PRION GLOBAL REPORT UPDATE JULY 17 2016
http://chronic-wasting-disease.blogspot.com/2016/07/chronic-wasting-disease-cwd-tse-prion.html
kind regards, terry
