OIE Reference Laboratory Reports
Activities in 2012
Name of disease (or topic) for which you
are a designated OIE Reference Laboratory:
Address of laboratory
Scrapie and Chronic Wasting Disease
Canadian Food Inspection Agency
Ottawa Laboratory-Fallowfield
3851 Fallowfield Rd, Ottawa, ON K2H8P9
Canada
Tel.:
613-228-6698
Fax:
613-221-6103
e-mail address:
aru.balachandran@inspection.gc.ca
website:
http://www.inspection.gc.ca/english/toce.shtml
Name (including Title) of Head of
Laboratory (Responsible Official):
Karen Jesset
Executive Director, Ontario Laboratory Network
Canadian Food Inspection Agency
Name(including Title and Position) of OIE
Reference Expert:
Dr. Aru Balachandran
Veterinary Science Advisor; Head, National and OIE
Reference Laboratory for Scrapie and CWD
Date of submission to the OIE
Annual reports of OIE Reference Centres, 2012
2013/01/25
1
OIE RL for « Scrapie and Chronic Wasting Disease (CWD) » – « Dr. Aru Balachandran » – « Canada »
ToR: To use, promote and disseminate diagnostic methods validated according to OIE
Standards
Disease
Test
For
Specificity
Total
Scrapie
IHC
Antigen
PrP Sc
1069
BioRad ELISA
VLA Hybrid & BioRad
TeSeE-Confirm WB
Antigen
PrP
Sc
7410
Antigen
PrP Sc
Genotyping sheep
Codon136,154,171
IHC
Antigen
PrP CWD
704
ELISA
VLA Hybrid & BioRad
TeSeE-Confirmatory
WB
Genotyping-Elk,
white-tailed deer
Antigen
PrP CWD
1316
Antigen
PrP CWD
80
CWD
97
2518
Codon132, 96
152
BSE
ELISA
Antigen
PrP BSE
AMR
IHC
Antigen
GFAP
4990
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
3.
No
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
5.
No
Did your laboratory produce vaccines?
Yes
6.
2
No
Did your laboratory supply vaccines to OIE Member Countries?
Annual reports of OIE Reference Centres, 2012
OIE RL for « Scrapie and Chronic Wasting Disease (CWD) » – « Dr. Aru Balachandran » – « Canada »
Yes
No
ToR: To develop, standardise and validate, according to OIE Standards, new procedures for
diagnosis and control of the designated pathogens or diseases
7.
Did your laboratory develop new diagnostic methods validated according to OIE Standards for the
designated pathogen or disease?
Yes
8.
No
Did your laboratory develop new vaccines according to OIE Standards for the designated pathogen
or disease?
Yes
Name of the new test or diagnostic
method or vaccine developed
No
Description and References (Publication, website, etc.)
Cell culture for the in vitro detection of
prion infectivity
The requirement for rapid and robust assays to detect of
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
of ulta-low levels of pathologic prion
protein
Current testing methods for transmissible spongiform
encephalopathies (TSEs) involve the detection of abnormal
prion protein (PrPTSE) by immunochemical methods as the
hallmark of infection. Novel technologies that combine
higher sensitivity detection with fast, robust, and costefficient 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
resistance/susceptibility
Susceptibility of cervids to CWD infection has been shown
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 »
Rodent bioassay for the quantification and
characterization of prion infectivity
The rodent bioassay platform continues to develop towards
characterising Canadian scrapie and CWD strain variability,
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
Member Country
seeking assistance
Date
(dd/mm)
No. samples received for
provision of diagnostic support
(i.e. from surveillance campaign)
No. samples received for
provision of confirmatory
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
Name of the OIE Member Country
receiving a technical consultancy
Republic of Korea
Republic of Korea
Japan
4
No
How the advice was
provided
Purpose
Dr. Min-Jeong Kim of the Prion Disease
Research Laboratory from Animal, Plant,
and Fisheries Quarantine and Inspection
Agency spent a week in the TSE lab
training in diagnostic test methodology
and result interpretation.
Dr. Aru Balachandran: Site visit to QIA
lab for consultation on collaborative
research underway.
Dr. Takashi Yokoyama, head of the Prion
Diseases Research Unit from the
National Institute of Animal Health and
Dr. Motohiro Horiuchi, of Hokkaido
University, presented their labs
Visit to OLF-CFIA
Canada
Visit to QIA, South Korea
Visit to OLF-CIFA
Canada
Annual reports of OIE Reference Centres, 2012
OIE RL for « Scrapie and Chronic Wasting Disease (CWD) » – « Dr. Aru Balachandran » – « Canada »
developments and novel approaches
with Prion diseases.
Dr. Ma Guiping, China's leading advisor
on TSEs in AQSIQ, visited Canada to
exchange technical knowledge on BSE
and CWD with CFIA laboratories and to
learn more about Canadian federal,
provincial and industry programs related
to these issues.
China
Drs. Jen-Chieh Chang and Jen-Huan Tsai
from the Animal Health Research
Institute, Taipei spent a week in our
laboratory for technical training on
various TSE diagnostic methodologies.
Taiwan
Visit to OLF-CFIA
Canada
Visit to OLF-CFIA
Canada
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
Title of the study
Investigating
cross-species
transmission of
CWD
Duration
On-going
No
Partners
(Institutions)
Purpose of the study
To characterize South
Korean and Canadian
chronic wasting
disease strains using
cervid and transgenic
mouse models.
Dr. Hyun-Joo Sohn,
Dr. Yoon-Hee Lee,
Dr. Min-Jeong Kim,
Dr. In-Soo Cho
:Veterinary
Researchers at
QIA, South Korea.
OIE Member Countries
involved other than
your country
Republic of 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
13.
No
Did your laboratory disseminate epizootiological data that had been processed and analysed?
Yes
14.
No
What method of dissemination of information is most often used by your laboratory?
(Indicate in the appropriate box the number by category)
a) ...... Articles published in peer-reviewed journals:
b) ..... International conferences:
6
3
Annual reports of OIE Reference Laboratories, 2012
5
OIE RL for « Scrapie and Chronic Wasting Disease (CWD) » – « Dr. Aru Balachandran » – « Canada »
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:
3
b) .........................................................................................Seminars:
c) .........................................................................................Hands-ontraining courses:
d) .........................................................................................Internships (>1 month):
Type of technical training
provided (a, b,c or d)
Country of origin of the expert(s)
provided with training
No. participants from the
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
contaminated meat products by H&E, and
GFAP IHC staining
SCC
Detection of abnormal prion protein for
BSE, Scrapie and CWD Diagnosis by Bio-
SCC
6
Annual reports of OIE Reference Centres, 2012
OIE RL for « Scrapie and Chronic Wasting Disease (CWD) » – « Dr. Aru Balachandran » – « Canada »
Rad ELISA
Genotyping Sheep for Scrapie
Suceptibility/Resistance by Real-Time PCR
SCC
Confirmation of prion protein specific for
scrapie and CWD using Bio-Rad’s TeSeE
Western Blot Kit
SCC
Immunohistochemical detection of prion
protein in animal TSEs: Scrapie in Sheep
and Goats
SCC
Immunohistochemical detection of prion
protein in animal TSEs: Chronic wasting
disease (CWD) in Deer and Elk
SCC
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
20.
No
Did your laboratory participate in scientific meetings on behalf of the OIE?
Yes
Title of event
No
Date
(mm/yy)
Location
Role (speaker, presenting
poster, short
communications)
Title of the work
presented
Prion 2012
05/12
Amsterdam,
Netherlands
Speaker
Chronic Wasting Disease
(CWD) in Canada: Update
on cross-species
transmission studies
Annual meeting of
the American
College of
Veterinary
Pathologists
12/12
Seattle,
Washington,
USA
Poster presentation
Characterization of
Canadian CWD isolates in
transgenic mice.
1st QIA Workshop
and Hands-on
training on CWD and
other prion diseases,
in collaboration with
the CFIA, Canada
and the NIAH, Japan
6/12
QIA, Republic
of Korea
Speaker
Annual reports of OIE Reference Laboratories, 2012
TSEs of small ruminants
and cervids in Canada
7
OIE RL for « Scrapie and Chronic Wasting Disease (CWD) » – « Dr. Aru Balachandran » – « Canada »
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
22.
No
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
23.
No
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
Reference Laboratories
Title of the project or contact
Scope
Investigating cross-species
transmission of CWD
To characterize South Korean and
Canadian chronic wasting disease strains
using cervid and transgenic mouse
models.
Animal, Plant and Fisheries
Quarantine and Inspection
Agency, Republic of Korea.
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
Purpose for inter-laboratory test comparisons1
No
No. participating
laboratories
Participating OIE Member
Countries
2
USDA, Pullman WA and Colorado
State Veterinary laboratory,
Fort Collins, CO
Validation of IHC testing for scrapie and CWD
USA
ToR: To place expert consultants at the disposal of the OIE
25.
8
Did your laboratory place expert consultants at the disposal of the OIE?
Annual reports of OIE Reference Centres, 2012
OIE RL for « Scrapie and Chronic Wasting Disease (CWD) » – « Dr. Aru Balachandran » – « Canada »
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.
PrPC with locked “component 3” metal coordination geometry – Altered properties of PK-resistant Cterminal 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.
Annual reports of OIE Reference Laboratories, 2012
9