Rabies
OIE Reference Laboratory Reports
Activities in 2010
Name of disease (or topic) for which you are a designated OIE Reference Laboratory: / RabiesAddress of laboratory/ / Canadian Food Inspection Agency
Centre of Expertise for Rabies, Ottawa Laboratory Fallowfield
3851 Fallowfield Road
Ottawa, Ontario
K2S 8P9 Canada
Tel.: / (+1-613) 228.66.98
Fax: / (+1-613) 228.66.69
e-mail address: /
website: / http://www.inspection.gc.ca/english/anima/disemala/rabrag/rabrage.shtml
Name of Head of Laboratory (Responsible Official): / Dr Lindsay Elmgren
Name of OIE Reference Expert: / Dr Alexander Wandeler until his retirement in September 2010.
Dr Christine Fehlner-Gardiner has been nominated to replace him. o be decided by OIE Biological Standards Commission in February 2011
Name of writer of this report
(if different from above): / Christine Fehlner-Gardiner
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
Numbers stated for direct immunofluorescence, cell culture inoculation and virus typing include samples tested at the CofE for Rabies, Ottawa Laboratory Fallowfield (OLF) as well as the sister rabies laboratory located at the CFIA Lethbridge Laboratory (ADRI).
Test / For / Specificity / TotalDirect immunofluorescence / Antigen / Lyssavirus / 4880
Indirect immunofluorescence using monoclonal antibodies / Antigen (virus variant identification) / Virus variant specific / 123
Cell culture inoculation test / Virus isolation / Lyssavirus / 3665
Immunohistochemistry / Antigen / Rabies virus / 9
RT-PCR
/ Genomic RNA (human samples only) / Lyssavirus / 5Competitive ELISA
/ Rabies antibody* / Rabies virus glycoprotein / 1050Virus neutralization
/ Rabies antibody* / Rabies virus glycoprotein / 630*Serology tests were carried out as part of collaborative research agreements to assess efficacy of oral vaccination programs for wildlife and to characterize immune response in target and non-target species (i.e., no export testing is done)
2. Production and distribution of diagnostic reagents
Serum of goats immunized with gradient-purified rabies ribonucleoprotein (RNP) is labelled with fluorescein isothiocyante. The conjugate produced at OLF is also used at ADRI Lethbridge. OLF and ADRI Lethbridge are the only two laboratories performing rabies diagnostics in Canada. All direct immunofluorescence tests for rabies are conducted using the OLF conjugate.
Rabbit antiserum raised against RNP is produced for use in immunohistochemistry.
Monoclonal antibodies (Mabs) to N-, P-, G-, and M-proteins of various Lyssaviruses are produced in cooperation with the laboratory Hybridoma unit. The Mabs are used for virus species and variant identification. A panel suitable to differentiate between the virus variants occurring in Western Canada is supplied to the Rabies Lab at the CFIA Lethbridge Laboratory.
The OLF molecular biology group in cooperation with the CofE for Rabies produces specific molecular probes for PCR and in situ hybridization for rabies virus variant identification and molecular epidemiology.
Reagents supplied to other OIE Member Countries
Fluorescent conjugate, Mabs and rabbit antiserum are provided free of charge to other laboratories provided a Materials Transfer Agreement is established. In 2010 the CofE for Rabies supplied rabbit antiserum to the Kimron Veterinary Institute, Israel, for use in immunohistochemistry tests for rabies. Anti-phosphoprotein Mabs were provided to Onderstepoort Veterinary Institute Rabies Laboratory, South Africa for research purposes. Purified and enzymatically labelled Mabs specific for glycoproteins of Mokola, Lagos bat, Duvenhage, canid and mongoose rabies viruses were also provided to OVI. Purified and biotinylated anti-nucleoprotein Mabs were provided to the OIE Reference Laboratory in the US Centers for Disease Control for evaluation for use in a Mab cocktail for their Direct Rapid Immunohistochemical Test for Rabies (DRIT).
Type of reagent / Amount supplied nationally(including for own use) / Amount supplied to other countries
Immunofluorescent conjugate / 30 ml / none in 2010
Rabbit antiserum / 5 ml / 1 mL
Monoclonal antibodies
/ 10 ml each of ~25 Mabs / 2 mL each of ~15 Mabs (supernant) 1 ml each of 12 purified and labelled MabsPart 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
None.
4. Preparation and supply of international reference standards for diagnostic tests or vaccines
None.
5. Research and development of new procedures for diagnosis and control
A real-time RT-PCR method for rapid testing of human samples for rabies was developed and validation of this method has been ongoing for the last two years with a view to incorporating this technique into routine human testing.
The laboratory is continuously improving its rabies variant identification capabilities. Variant identification is conducted on all rabies positive submissions.
The laboratory previously collaborated with industy, academia and the Ontario Provincial Government in the development of a human adenovirus rabies-glycoprotein recombinant oral vaccine (ONRAB®). This new product is presently applied in wildlife rabies control projects in the provinces of Ontario, Quebec and New Brunswick. The CofE for Rabies participates in monitoring these field trials through rabies serology and determinations of vaccine stability. The C of E is continuing its research collaboration with the Ontario Ministry of Natural Resources examining the kinetics and duration of immune response of target species to the ONRAB® vaccine.
6. Collection, analysis and dissemination of epizootiological data relevant to international disease control
The CofE for Rabies continues to monitor a raccoon variant rabies outbreak in a cooperative research project with Quebec authorities.
An OLF Mab panel is used at the Onderstepoort OIE laboratory in South Africa for discriminating between Mokola, Lagos bat, canid and mongoose virus infections. In addition, the C of E is engaged with the Onderstepoort laboratory in the development of ELISAs for use in serosurveillance. The C of E has provided training and reagents to Onderstepoort personnel for this project (see questions #2 and #8).
7. Provision of consultant expertise to OIE or to OIE Members
The CofE for Rabies participated in a Technical Exchange Program with the OIE Rabies Reference Laboratory at the Onderstepoort Veterinary Insititue (OVI), South Africa, for the development of competitive ELISAs for use in serological surveys of Mokola, Lagos bat, canid and mongoose virus infections in South Africa. This project was funded by the Canadian International Development Agency (CIDA), began in 2009 and continued into 2010.
The C of E provided advice to collaborators at the Kyrgyz Research Institute of Livestock, Veterinary and Pastures for the ISTC-funded project “Monitoring of the epizootiological conditions and prevention of occurrence of rabies of people and animals”.
Dr. Fehlner-Gardiner continued to serve as an invited particpant in the USDA-APHIS Wildlife Services National Rabies Management Planning Meeting and as the CFIA representative at the North American Rabies Management Meetings.
8. Provision of scientific and technical training to personnel from other OIE Members
As part of the CIDA-funded project described above, the CofE for Rabies hosted one PhD candidate (2 months) and two professional staff (3 weeks) from the Onderstepoort Veterinary Insitute in Pretoria, South Africa. Personnel were trained in antibody purification and labelling techniques, ribonucleoprotein purification, molecular genetics techniques and software analysis tools, quality assurance, and diagnostic testing procedures.
9. Provision of diagnostic testing facilities to other OIE Members
None.
10. Organisation of international scientific meetings on behalf of OIE or other international bodies
None.
11. Participation in international scientific collaborative studies
In 2010, the C of E collaborated in a study with USDA-APHIS Wildlife Services, the Quebec Ministry of Natural Resources and the New York State Department of Health Rabies Laboratory, evaluating the field efficacy of ONRAB® and another recombinant oral vaccine for wildlife currently licensed for use in the USA. The C of E carried out the serological analyses of raccoon and skunk sera obtained from areas in Quebec and Vermont where the two vaccines were distributed. The project is currently in the data analysis stage.
12. 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
Hamill PH, Kush J, Nadin-Davis AS, Wandeler A, Fehlner-Gardiner C. 2010. Rabies in Canada – 2009. 21st International Conference on Rabies in the Americas. Guadalajara, Mexico.
Hanlon CA, Wong S, Gordon C, Niezgoda M, Fehlner-Gardiner C, Wandeler A, Moore S. 2010. Measures of rabies immunity and optimal practices in rabies prevention. 21st International Conference on Rabies in the Americas. Guadalajara, Mexico.
Fehlner-Gardiner C. 2010. Development and validation of a competitive ELISA for wildlife serology – Lessons learned. Kansas State University Rabies Laboratory Workshop on Rabies Immunology. Manhattan, KS.
¡ Scientific publications in peer-reviewed journals
Nadin-Davis SA, Fehlner-Gardiner C, Sheen M, Wandeler AI. 2010. Characterization of a panel of anti-phosphoprotein monoclonal antibodies generated against the raccoon strain of rabies virus. Virus Research 152:126-36.
Sobey KG, Rosatte R, Bachmann P, Buchanan T, Bruce L, Donovan D, Brown L, Davies JC, Fehlner-Gardiner C, Wandeler A. 2010. Field evaluation of an inactivated vaccine to control raccoon rabies in Ontario, Canada. Jounal of Wildlife Diseases 46:818-31.
Nadin-Davis SA, Feng Y, Mousse D, Wandeler AI, Aris-Brosou S. 2010 Spatial and temporal dynamics of rabies virus variants in big brown bat populations across Canada: footprints of an emerging zoonosis. Molecular Ecology 19: 2120-2136.
Brown LJ, Rosatte RC, Fehlner-Gardiner C, Knowles MK, Bachmann P, Davies JC, Wandeler A, Sobey K, Donovan D. 2010. Immunogenicity and efficacy of two rabies vaccines in wild-caught, captive raccoons. Jounal of Wildlife Diseases In press.
¡ Other communications
Fehlner-Gardiner C. 2010. Rabies in Canada and Development and validation of a competitive ELISA for wildlife serology. Seminars given at the Onderstepoort Veterinary Institute. Pretoria, South Africa.
13. Inscription of diagnostic kits on the OIE Register
i) Did you participate in expert panels for the validation of candidate kits for inscription on the OIE Register? If yes, for which kits?
No.
ii) Did you submit to the OIE candidate kits for inscription on the OIE Register? If yes, for which kits?
No.
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Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010 5