OIE Reference Laboratory Reports
Activities in 2011
Name of disease (or topic) for
which you are a designated OIE
Reference Laboratory:
Address of laboratory:
Highly pathogenic avian influenza and low
pathogenic avian influenza (poultry)
Canadian Food Inspection Agency
National Centre for Foreign Animal Disease
1015 Arlington Street, Winnipeg, Manitoba
CANADA R3E 3M4
Tel.:
1-204-789-2013
Fax:
1-204-789-2038
e-mail address:
John.Pasick@inspection.gc.ca
website:
Name (including Title and
Position) of Head of Laboratory
(Responsible Official):
Name(including Title and
Position) of OIE Reference
Expert:
Name (including Title and
Position) of writer of this report
(if different from above):
Dr Soren Alexandersen,
Executive Director,
National Centre for Animal Diseases
Dr John Pasick, Head,
Classical Swine Fever &
Avian Diseases Section
National Centre for
Foreign Animal Disease
Highly pathogenic avian influenza and low pathogenic avian influenza (poultry)
Part I: Summary of general activities related to the disease
1.
Test(s) in use/or available for the specified disease/topic at your laboratory
Test
For
Specificity
Total
cELISA (influenza A)
Antibody
Group
9548
AGID
Antibody
Group
755
HI (H3, H5 & H7)
Antibody
Type
505
HI (other)
Antibody
Type
78 (78 × 16 subtypes)
Real-Time RT-PCR (influenza
A)
Viral nucleic acid
Group
5069
Real-Time RT-PCR (H5 &
H7)
Viral nucleic acid
HA subtype
1900
Virus isolation (embryonating
chicken eggs)
Virus
Group
538
HA Typing
Virus characterization
HA subtype
68
NA Typing
Virus characterization
NA subtype
832
Nucleic acid sequencing &
molecular pathotyping
Virus characterization
Genome sequence HA0
cleavage site
18
IVPI
Virus pathogenicity
2.
4
Production and distribution of diagnostic reagents
Type of reagent
Amount supplied nationally
(including for own use)
Amount supplied to
other countries
H5 reference antiserum
~500 mL produced primarily for in house use
14 mL
In vitro transcribed RNA controls
for real-time RT-PCR assays
(matrix, H5 and H7)
Matrix: 9.0 ml (90 vials @ 100 µl/vial)
H5: 1.6 ml (16 vials @ 100 µl/vial)
H7: 1.7 ml (17 vials @ 100 µl/vial)
Matrix: 2 × 100 µl
H5: 2 × 100 µl
H7: 2 × 100 µl
NP, H5 and H7 monoclonal
antibodies
NP monoclonal Ab: 400 µL (2 vials @ 200 µL/vial)
Positive antisera to the 16 HA
subtypes and 9 NA subtypes
~4.8 litres produced
16 ml
Recombinant Influenza A virus
NP protein for use in enzymelinked immunosorbent assays
300 µL (3 vials @ 100 µl/vial)
None
Real-time RT-PCR proficiency
panels
33 panels
4 panels (20
samples/panel)
Influenza A competitive ELISA
proficiency panels
10 mL (1 panel @ 10 mL/panel)
None
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Annual reports of OIE Reference Centres, 2011
Highly pathogenic avian influenza and low pathogenic avian influenza (poultry)
Part II: Activities specifically related to the mandate
of OIE Reference Laboratories
3.
International harmonisation and standardisation of methods for diagnostic testing or the
production and testing of vaccines
a)
Establishment and maintenance of a network with other OIE Reference Laboratories
designated for the same pathogen or disease and organisation of regular inter-laboratory
proficiency testing to ensure comparability of results
 Participated in an avian influenza proficiency test panel which focused on molecular and serological
subtyping assays. The panel was produced by the OFFLU PT technical working group. The goal of the
exercise was to standardize diagnostic testing for avian influenza virus through participation in a worldwide proficiency testing system for international reference laboratories.
 Participated in avian influenza and avian paramyxovirus real-time RT-PCR proficiency test with the
USDA.
b)
Organisation of inter-laboratory proficiency testing with laboratories other than OIE
Reference Laboratories for the same pathogens and diseases to ensure equivalence of
results
 Participation in a regional working group involving the national laboratories of the United States, Canada
and Mexico whose objective is to harmonize avian influenza diagnostic test methods. As part of this
working group, our laboratory produced an H5/H7 hemagglutination-inhibition assay panel which was
sent out to the participating laboratories in the USA and Mexico in September 2011.
4.
Preparation and supply of international reference standards for diagnostic tests or vaccines
A test lot (28 × 0.5 ml) of candidate H5 Reference Standard Serum along with its homolgous antigen was sent to
the HPAI Reference Laboratory in Weybridge. These will be distributed to other OIE HPAI Reference Laboratories
for a second round of evaluation.
5.
Research and development of new procedures for diagnosis and control
Validation of a competitive ELISA specific for H5 subtype antibodies continues. Validation is currently in Stage
III. A manuscript describing assay development and validation has been published.
Baculovirus expressed HA and NA antigens are being utilized in the development of a multiplex suspension
microarray (Luminex).
Development of RT-PCR-Pyrosequencing coupled assays to rapidly genotype the HA0 cleavage site of H5 and H7
avian influenza viruses continues. Currently in stage I validation.
Development of DNA microarray-based assays for determining the HA and NA subtype of avian influenza virus
isolates (collaborative project with ADRI, Lethbridge, AB). Stage II validation has been completed. A manuscript
is in preparation.
Identification and characterization of virulence determinants of avian influenza viruses using reverse genetics.
6.
Collection, analysis and dissemination of epizootiological data relevant to international disease
control
The Canadian Food Inspection Agency (CFIA) is conducting the Canadian Notifiable Avian Influenza (NAI)
Surveillance System (CanNAISS) in poultry populations (i.e., chickens, turkeys, ducks, geese and specialty birds)
in Canada. The goal of CanNAISS is to assess if NAI is circulating in commercial chicken, turkey, ducks, geese
and specialty bird farms. Active surveillance with on-farm sampling to test for NAI is ongoing in two components:
Annual reports of OIE Reference Centres, 2011
3
Highly pathogenic avian influenza and low pathogenic avian influenza (poultry)
The pre-slaughter surveillance component targets healthy poultry flocks. The selection of farms is based on
slaughter schedules from federally inspected processing plants in Canada.
The hatchery supply flock surveillance component also targets healthy poultry flocks. Flocks supplying hatching
eggs to health monitored hatcheries, as defined in Section 79 of the Health of Animals Regulations, are sampled as
they approach the end-of-production.
NCFAD supports this surveillance activity by doing all the diagnostic testing required.
In 2011 NCFAD screened approximately 6640 birds using the cELISA and did secondary testing (HI) on
77 samples for CanNAISS. No notifiable AI was detected in 2011.
Survey of influenza viruses in wild aquatic and migratory birds continued in 2011.
7.
Maintenance of a system of quality assurance, biosafety and biosecurity relevant to the
pathogen and the disease concerned
ISO 17025 accredited for diagnostics and research. We have Animal Pathogen Containment levels 2, 3, 3+ and 4
(BSL 2, 3, 3+ and 4). Our laboratories and animal cubicles are certified annually by the Canadian Food Inspection
Agency and the Public Health Agency of Canada to meet the above.
8.
Provision of consultant expertise to OIE or to OIE Member Countries
A member of our laboratory represented OFFLU at the WHO Meeting of the Technical Group on PCR Diagnosis
of Influenza Virus Infections, Geneva, 14-15 June, 2011.
Reviewed the chapter on Avian Influenza for the 7th edition of the OIE Manual of Diagnostic Tests and Vaccines
for Terrestrial Animals.
9.
Provision of scientific and technical training to personnel from other OIE Member Countries
Hosted a 3-day workshop in December 2011 which focused on the molecular analysis of animal influenza A
viruses. This workshop was sponsored by PROCINORTE and concentrated on influenza A virus whole genome
sequencing and analysis. Three scientists from the USA and three scientists from Mexico participated in the
workshop.
Work continued on an OIE twinning project entitled “Technical Support to the LNDV for the Diagnosis and
Control of Avian Influenza and Newcastle Disease” with the National Diagnostic Laboratory in Bogota, Colombia
(LNDV-ICA-Colombia). Diagnostic reagents which included avian influenza subtype reference antisera, real-time
RT-PCR primers, probes and controls and real-time RT-PCR proficiency panels were shipped to LNDV-ICAColombia.
10. Provision of diagnostic testing facilities to other OIE Member Countries
None in 2011.
11. Organisation of international scientific meetings on behalf of OIE or other international bodies
None in 2011.
12. Participation in international scientific collaborative studies
None in 2011.
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Annual reports of OIE Reference Centres, 2011
Highly pathogenic avian influenza and low pathogenic avian influenza (poultry)
13. Publication and dissemination of information relevant to the work of OIE (including list of
scientific publications, internet publishing activities, presentations at international conferences)

Presentations at international conferences and meetings
Berhane Y, Neufeld J, Kehler H, Leith M, Suderman M, Pasick J. Pre-exposure of Canada geese to low pathogenic
avian influenza H1N1 virus protects against lethal H5N1 infection. 5 th Vaccine and ISV Annual Global
Conference. Seattle, Washington, USA, 2-4 October 2011.

Scientific publications in peer-reviewed journals
Parmley EJ, Soos C, Breault A, Fortin M, Jenkins E, Kibenge F, King R, McAloney K, Pasick J, Pryor SP,
Robinson J, Rodrigue J, Leighton FA. 2011. Detection of low pathogenic avian influenza viruses in wild ducks
from Canada: comparison of two sampling methods. J Wildlife Dis 47: 466-470.
Yang M, Clavijo A, Pasick J, Salo T, Wang Z, Zhao Y, Zheng D, Berhane Y. 2011. Serologic detection of avian
influenza H5 antibodies using a competitive enzyme-linked immunosorbent assay (ELISA). J Veterinary Medicine
and Animal Health 3: 56-61.

Other communications
None in 2011.
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Annual reports of OIE Reference Centres, 2011
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