Highly pathogenic avian influenza and low pathogenic avian influenza (poultry)

OIE Reference Laboratory Reports

Activities in 2011

Name of disease (or topic) for which you are a designated OIE Reference Laboratory: / Highly pathogenic avian influenza and low pathogenic avian influenza (poultry)
Address of laboratory: / Canadian Food Inspection Agency
National Centre for Foreign Animal Disease
1015 Arlington Street, Winnipeg, Manitoba
CanadaR3E 3M4
Tel.: / 1-204-789-2013
Fax: / 1-204-789-2038
e-mail address: /
website:
Name (including Title and Position) of Head of Laboratory (Responsible Official): / Dr SorenAlexandersen,
Executive Director,
National Centre for Animal Diseases
Name(including Title and Position) of OIE Reference Expert: / Dr John Pasick, Head,
Classical Swine Fever &
Avian Diseases Section
National Centre for
Foreign Animal Disease
Name (including Title and Position) of writer of this report
(if different from above):

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 (embryonatingchicken 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 / 4

2.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 enzyme-linked 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

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 world-wide 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:

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 2011NCFADscreened approximately 6640birds using the cELISA and did secondary testing (HI) on 77samples 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-ICA-Colombia.

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.

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. 5th 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 immunosorbentassay (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, 20111