OIE Reference Laboratory Reports Activities in 2012

Name of disease (or topic) for which you are a designated OIE Reference Laboratory:

Address of laboratory

Tel.:

Fax: e-mail address: website:

Name (including Title) of Head of Laboratory (Responsible Official):

Name(including Title and Position) of OIE Reference Expert:

Date of submission to the OIE

ToR: To use, promote and disseminate diagnostic methods validated according to OIE Standards

Diseas Test For Specificity Total e Scrapie IHC Antigen PrP Sc 1069

Annual reports of OIE Reference Centres, 2012 1 OIE RL for « Scrapie and Chronic Wasting Disease (CWD) » – « Dr. Aru Balachandran » – « Canada »

BioRad ELISA Antigen PrP Sc 7410 VLA Hybrid & BioRad TeSeE-Confirm WB Antigen PrP Sc 97 Genotyping sheep Codon136,154,171 2518 CWD IHC Antigen PrP CWD 704 ELISA Antigen PrP CWD 1316 VLA Hybrid & BioRad TeSeE-Confirmatory WB Antigen PrP CWD 80 Genotyping-Elk, white-tailed deer Codon132, 96 152 BSE ELISA Antigen PrP BSE 4990 AMR IHC Antigen GFAP 316

ToR: To develop reference material in accordance with OIE requirements, and implement and promote the application of OIE Standards. To store and distribute to national laboratories biological reference products and any other reagents used in the diagnosis and control of the designated pathogens or disease.

2. Did your laboratory produce or store imported standard reference reagents officially recognised by the OIE or other international bodies?

Yes No

3. Did your laboratory supply standard reference reagents to OIE Member Countries?

Yes No

While most diagnostic reagents required for scrapie and CWD testing are commercially available, we continue to develop and maintain an archive of TSE-positive and negative reference materials for use in test development, validation and proficiency testing programs. These materials are used nationally within our reference and network laboratories and have been provided to countries in North and South America and Asia who are seeking to develop TSE testing programs or validate existing programs.

4. Did your laboratory produce diagnostic reagents other than the OIE-approved standard reference reagents?

Yes No

5. Did your laboratory produce vaccines?

Yes No

6. Did your laboratory supply vaccines to OIE Member Countries?

Yes No

ToR: To develop, standardise and validate, according to OIE Standards, new procedures for diagnosis and control of the designated pathogens or diseases

2 Annual reports of OIE Reference Centres, 2012 OIE RL for « Scrapie and Chronic Wasting Disease (CWD) » – « Dr. Aru Balachandran » – « Canada »

7. Did your laboratory develop new diagnostic methods validated according to OIE Standards for the designated pathogen or disease?

Yes No

8. Did your laboratory develop new vaccines according to OIE Standards for the designated pathogen or disease?

Yes No

Name of the new test or diagnostic Description and References (Publication, website, etc.) method or vaccine developed

Cell culture for the in vitro detection of The requirement for rapid and robust assays to detect of prion infectivity low levels of prion proteins in a variety of sample matricies continues. Cell culture based systems have recently demonstrated utility in the detection of scrapie and CWD, and we continue to refine analytical sensitivity towards broader diagnostic and research applications.

Evaluation of immuno-PCR for the detection Current testing methods for transmissible spongiform of ulta-low levels of pathologic prion encephalopathies (TSEs) involve the detection of abnormal protein prion protein (PrPTSE) by immunochemical methods as the hallmark of infection. Novel technologies that combine higher sensitivity detection with fast, robust, and cost- efficient protocols hold promise for future diagnostic and research applications. Work is in progress to increase the analytical sensitivity of immuno-PCR relative to currently approved test methods and to investigate the applicability for pre-clinical diagnosis and environmental testing.

Cervid genotyping for CWD Susceptibility of cervids to CWD infection has been shown resistance/susceptibility to be associated with polymorphisms in the host prion protein gene. Work is in progress to validate recently developed high throughput methods to detect the allelic variants of elk, white-tailed deer at codons 132, and 96 respectively. This information may be used as a component of regulatory control and eradication programs. Measures of sensitivity and specificity do not apply as per diagnostic testing of a disease/infectious agents since this test provides the genotype of a particular animal and not infected/non-infected status. The method used to detect these allelic variants has undergone partial Level 1 formal validation in accordance with the OIE Validation Template (calibration, repeatability studies).

Annual reports of OIE Reference Laboratories, 2012 3 OIE RL for « Scrapie and Chronic Wasting Disease (CWD) » – « Dr. Aru Balachandran » – « Canada »

The rodent bioassay platform continues to develop towards Rodent bioassay for the quantification and characterising Canadian scrapie and CWD strain variability, characterization of prion infectivity investigating species barriers to infectivity and determining infectivity titres from a variety of matrices. Diagnostic utility has been demonstrated in differentiating between CWD, atypical and classical scrapie and BSE in field samples. The ability to quantify infectivity enables the development and validation of novel ultrasensitive tests such as PMCA, immuno-PCR, RT-QuIC and cell culture.

Contact : Animal Health Research and Partnership Division, Canadian Food Inspection Agency Floor 1, Room 314, 1400, Merivale Road, Tower 1, Ottawa , Ontario, K1A 0Y0 Canada,

ToR: To provide diagnostic testing facilities, and, where appropriate, scientific and technical advice on disease control measures to OIE Member Countries

9. Did your laboratory carry out diagnostic testing for other OIE Member Countries?

Yes No

Name of OIE Date No. samples received for No. samples received for Member Country (dd/mm) provision of diagnostic support provision of confirmatory seeking assistance (i.e. from surveillance campaign) diagnoses

Brazil 12/10 92 0

10. Did your laboratory provide expert advice in technical consultancies on the request of an OIE Member Country?

Yes No

Name of the OIE Member Country How the advice was Purpose receiving a technical consultancy provided

Republic of Korea Dr. Min-Jeong Kim of the Prion Disease Visit to OLF-CFIA Research Laboratory from Animal, Plant, Canada and Fisheries Quarantine and Inspection Agency spent a week in the TSE lab training in diagnostic test methodology and result interpretation.

Republic of Korea Dr. Aru Balachandran: Site visit to QIA Visit to QIA, South Korea lab for consultation on collaborative research underway.

Japan Dr. Takashi Yokoyama, head of the Prion Visit to OLF-CIFA Diseases Research Unit from the Canada National Institute of Animal Health and Dr. Motohiro Horiuchi, of Hokkaido

4 Annual reports of OIE Reference Centres, 2012 OIE RL for « Scrapie and Chronic Wasting Disease (CWD) » – « Dr. Aru Balachandran » – « Canada »

University, presented their labs developments and novel approaches with Prion diseases. Dr. Ma Guiping, China's leading advisor China Visit to OLF-CFIA on TSEs in AQSIQ, visited Canada to exchange technical knowledge on BSE Canada and CWD with CFIA laboratories and to learn more about Canadian federal, provincial and industry programs related to these issues.

Drs. Jen-Chieh Chang and Jen-Huan Tsai Taiwan Visit to OLF-CFIA from the Animal Health Research Institute, Taipei spent a week in our Canada laboratory for technical training on various TSE diagnostic methodologies.

ToR: To carry out and/or coordinate scientific and technical studies in collaboration with other laboratories, centres or organisations

11. Did your laboratory participate in international scientific studies in collaboration with OIE Member Countries other than the own?

Yes No

OIE Member Countries Partners Title of the study Duration Purpose of the study involved other than (Institutions) your country

Investigating On-going To characterize South Dr. Hyun-Joo Sohn, Republic of Korea cross-species Korean and Canadian Dr. Yoon-Hee Lee, transmission of chronic wasting Dr. Min-Jeong Kim, CWD disease strains using Dr. In-Soo Cho cervid and transgenic :Veterinary mouse models. Researchers at QIA, South Korea.

ToR: To collect, process, analyse, publish and disseminate epizootiological data relevant to the designated pathogens or diseases

12. Did your Laboratory collect epizootiological data relevant to international disease control?

Yes No

13. Did your laboratory disseminate epizootiological data that had been processed and analysed?

Yes No

14. What method of dissemination of information is most often used by your laboratory?

(Indicate in the appropriate box the number by category)

Annual reports of OIE Reference Laboratories, 2012 5 OIE RL for « Scrapie and Chronic Wasting Disease (CWD) » – « Dr. Aru Balachandran » – « Canada »

a)...... Articles published in peer-reviewed journals: 6 b)...... International conferences: 3 c)...... National conferences: X d)...... Other:

ToR: To provide scientific and technical training for personnel from OIE Member Countries

To recommend the prescribed and alternative tests or vaccines as OIE Standards

15. Did your laboratory provide scientific and technical training to laboratory personnel from other OIE Member Countries?

Yes No

If the answer is yes, please provide the total number of trained persons for each of the following categories:

a) Technical visits:...... b) Seminars:...... c) Hands-ontraining courses:...... d) Internships (>1 month):......

Type of technical training Country of origin of the expert(s) No. participants from the provided (a, b,c or d) provided with training corresponding country

A Taiwan 2

A Republic of Korea 1

ToR: To maintain a system of quality assurance, biosafety and biosecurity relevant for the pathogen and the disease concerned

16. Does your laboratory have a Quality Management System certified according to an International Standard?

Yes No

Quality management system adopted

ISO 17025

17. Is your laboratory accredited by an international accreditation body?

Yes No

Test for which your laboratory is accredited Accreditation body

Detection of spinal cord in beef SCC contaminated meat products by H&E, and GFAP IHC staining

6 Annual reports of OIE Reference Centres, 2012 OIE RL for « Scrapie and Chronic Wasting Disease (CWD) » – « Dr. Aru Balachandran » – « Canada »

Detection of abnormal prion protein for SCC BSE, Scrapie and CWD Diagnosis by Bio-Rad ELISA

Genotyping Sheep for Scrapie SCC Suceptibility/Resistance by Real-Time PCR

Confirmation of prion protein specific for SCC scrapie and CWD using Bio-Rad’s TeSeE Western Blot Kit

Immunohistochemical detection of prion SCC protein in animal TSEs: Scrapie in Sheep and Goats

Immunohistochemical detection of prion SCC protein in animal TSEs: Chronic wasting disease (CWD) in Deer and Elk

18. Does your laboratory maintain a “biorisk management system” for the pathogen and the disease concerned? (See Manual of Diagnostic Tests and Vaccines for Terrestrial Animals 2012, Chapter 1.1.3 or Manual of Diagnostic Tests for Aquatic Animals 2012, Chapter 1.1.1)

Yes No

ToR: To organise and participate in scientific meetings on behalf of the OIE

19. Did your laboratory organise scientific meetings on behalf of the OIE?

Yes No

20. Did your laboratory participate in scientific meetings on behalf of the OIE?

Yes No

Title of event Date Location Role (speaker, presenting Title of the work (mm/yy) poster, short presented communications)

Prion 2012 05/12 Amsterdam, Speaker Chronic Wasting Disease Netherlands (CWD) in Canada: Update on cross-species transmission studies

Annual meeting of 12/12 Seattle, Poster presentation Characterization of the American Washington, Canadian CWD isolates in College of USA transgenic mice. Veterinary Pathologists

1st QIA Workshop 6/12 QIA, Republic Speaker TSEs of small ruminants and Hands-on of Korea and cervids in Canada training on CWD and other prion diseases, in

Annual reports of OIE Reference Laboratories, 2012 7 OIE RL for « Scrapie and Chronic Wasting Disease (CWD) » – « Dr. Aru Balachandran » – « Canada »

collaboration with the CFIA, Canada and the NIAH, Japan

ToR: To establish and maintain a network with other OIE Reference Laboratories designated for the same pathogen or disease and organise regular inter-laboratory proficiency testing to ensure comparability of results

21. Did your laboratory exchange information with other OIE Reference Laboratories designated for the same pathogen or disease?

Yes No

22. Was your laboratory involved in maintaining a network with OIE Reference Laboratories designated for the same pathogen or disease by organising or participating in proficiency tests?

Yes No

23. Did your laboratory collaborate with other OIE Reference Laboratories for the same disease on scientific research projects for the diagnosis or control of the pathogen of interest?

Yes No

Name(s) of relevant OIE Title of the project or contact Scope Reference Laboratories

Investigating cross-species To characterize South Korean and Animal, Plant and Fisheries transmission of CWD Canadian chronic wasting disease strains Quarantine and Inspection using cervid and transgenic mouse Agency, Republic of Korea. models.

ToR: To organise inter-laboratory proficiency testing with laboratories other than OIE Reference Laboratories for the same pathogens and diseases to ensure equivalence of results.

24. Did your laboratory organise or participate in inter-laboratory proficiency tests with laboratories other than OIE Reference Laboratories for the same disease?

Yes No

No. participating Participating OIE Member Purpose for inter-laboratory test comparisons1 laboratories Countries

Validation of IHC testing for scrapie and CWD USDA, Pullman WA and Colorado State Veterinary laboratory, 2 Fort Collins, CO USA

ToR: To place expert consultants at the disposal of the OIE

1

8 Annual reports of OIE Reference Centres, 2012 OIE RL for « Scrapie and Chronic Wasting Disease (CWD) » – « Dr. Aru Balachandran » – « Canada »

25. Did your laboratory place expert consultants at the disposal of the OIE?

Yes No

Scientific Publications in peer-reviewed journals

Biodegradation of specified risk material and fate of scrapie prions in compost. Xu S, Reuter T, Gilroyed BH, Dudas S, Graham C, Neumann NF, Balachandran A, Czub S, Belosevic M, Leonard JJ, McAllister TA. J Environ Sci Health A Tox Hazard Subst Environ Eng. 2013;48(1):26-36.

Strain Characterization of the Korean CWD Cases in 2001 and 2004. Lee YH, Sohn HJ, Kim MJ, Kim HJ, Lee WY, Yun EI, Tark DS, Cho IS, Balachandran A. J Vet Med Sci. 2012 Aug 28

Diagnostic accuracy of rectal mucosa biopsy testing for chronic wasting disease within white-tailed deer (Odocoileus virginianus) herds in North America: effects of age, sex, polymorphism at PRNP codon 96, and disease progression. Thomsen BV, Schneider DA, O'Rourke KI, Gidlewski T, McLane J, Allen RW, McIsaac AA, Mitchell GB, Keane DP, Spraker TR, Balachandran A. J Vet Diagn Invest. 2012 Sep;24(5):878-87

Experimental oral transmission of chronic wasting disease to reindeer (Rangifer tarandus tarandus). Mitchell GB, Sigurdson CJ, O'Rourke KI, Algire J, Harrington NP, Walther I, Spraker TR, Balachandran A. PLoS One. 2012;7(6):e39055.

Rapid detection of CWD PrP: comparison of tests designed for the detection of BSE or scrapie. Blasche T, Schenck EV, Balachandran A, Miller MW, Langenberg J, Frölich K, Steinbach F. Transbound Emerg Dis. 2012 Oct;59(5):405-15

Inactivation of template-directed misfolding of infectious prion protein by ozone. Ding N, Neumann NF, Price LM, Braithwaite SL, Balachandran A, Belosevic M, El-Din MG. Appl Environ Microbiol. 2012 Feb;78(3):613-20

Presentations at international conferences or meetings

Chronic Wasting Disease (CWD) in Canada: Update on Cross-species transmission studies. Balachandran A, Mitchell GB, Sigurdson CJ, O’Rourke KI, Algire J, Harrington NP, Walther I, Spraker TR. Prion 2012, Amsterdam, The Netherlands, 2012.

Characterization of Canadian CWD isolates in transgenic mice. Mitchell G, Staskevicius A, Shaffer P, Foster S, Ghazi D, Yogasingam N, Walther I and Balachandran A. Annual meeting of the American College of Veterinary Pathologists, Seattle, Washington, USA, 2012.

Properties of geometry-locked PrPs and Shadoo protein in health and disease. Lau A, Daude N, McDonald A, Aglietti R, Mays CE, Walter ED, Visconte M, Wohlgemuth S, Yong J, Gapeshina H, Grams J, Balachandran A, Schmitt-Ulms G, Pushie MJ, George GN, Carlson GA, Millhauser G, Westaway D. Prion 2012, Amsterdam, The Netherlands, 2012.

PrP C with locked “component 3” metal coordination geometry – Altered properties of PK-resistant C-

Annual reports of OIE Reference Laboratories, 2012 9 OIE RL for « Scrapie and Chronic Wasting Disease (CWD) » – « Dr. Aru Balachandran » – « Canada »

terminal fragments and species-barriers to infection. Lau A, Mays C, Wohlgemuth S, Daude N, Yang J, Gapeshina H, Balachandran A, Millhauser G, Westaway D. Prion 2012, Amsterdam, The Netherlands, 2012.

10 Annual reports of OIE Reference Centres, 2012