Biotechnology-Based Diagnosis of Infectious
Diseases in Veterinary Medicine
Prof. Sándor Belák, Dr Peter Thorén & Dr. Karl Ståhl
National Veterinary Institute, Department of Virology
Travvägen 22, 751 89 Uppsala, SWEDEN
Tel.: (0046 18 674 000), Fax: (0046 18 674 669)
sandor.belak@sva.se; www.sva.se/oie-cc
Summary of activities specifically related to the mandate of
OIE Collaborating Centres
1.
Activities as a centre of research, expertise, standardisation and dissemination of techniques
within the remit of the mandate given by the OIE
Research in the frame of EU projects
The Collaborating Centre is currently conducting research and training in nine EU projects that include tasks on
the improvement of molecular and “on site” diagnostics, development of new methods for improved food safety,
as well as the international harmonisation and standardisation of the diagnostic procedures. The work is performed
at the Joint Research and Development Division of Departments of Virology of the National Veterinary Institute
(SVA, http://www.sva.se/) and of the Swedish University of Agricultural Sciences (SLU, http://www.slu.se/), in
collaboration with the other sections of the Department of Virology, SVA, first of all the Section of Molecular
Diagnostics. Further local collaborators are the Department of Pathology, SVA, Svanova Biotech AB, Olink AB,
the Section of Clinical Virology, Department of Medical Sciences, as well as the Department of Genetics and
Pathology/Molecular Medicine, Uppsala University, Uppsala, Sweden. Internationally, the collaborators are the
large consortia of the nine ongoing EU projects, representing a very wide network of animal health issues over
Europe and other continents (see below). The collaboration is active also with our sister Collaborating Centre at
IAEA in Vienna: the “OIE Collaborating Centre for ELISA and Molecular Technologies in Animal Disease
Diagnosis”.
Recently closed project, results are continuously obtained, dissemination is ongoing, website is regularly
updated:
QLK2-CT-2000-00486: Development, standardisation and harmonisation of novel multiplex nucleic acid tests for
the detection of economically important viruses of farm animals. Acronym: MULTIPLEX PCR (2000-2004).
Web address: http://www.multiplex-eu.org/
Role: Co-ordinator
Ongoing projects:
SSP3-513 645: New and emerging technologies: improved laboratory and on-site detection of OIE List A viruses
in animals and animal products. Acronym: LAB-ON-SITE (2004-2008).
Web address: http://www.labonsite.com/
Role: Co-ordinator
SSP1- 501599: Epidemiology and control of classical swine fever (CSF) in wild boar and potential use of a newly
developed live marker vaccine. Acronym: CSFVACCINE & WILDBOAR (2004-2007).
Web address: http://www.csfvaccine.org/
Role: Partner
Food-CT-2004-506122: Network for Prevention and Control of Zoonoses. Acronym: MED-VET-NET (20042009).
Web address: http://www.medvetnet.com/
Role: Partner
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Biotechnology-Based Diagnosis of Infectious Diseases in Veterinary Medicine
Food-CT-2005-007081: Development of cost-effective control and prevention strategies for emerging and future
foodborne pathogenic microorganisms throughout the food chain. Acronym: PATHOGEN COMBAT (20052010).
Web address: http://www.pathogencombat.com:80/
Role: Partner (SLU)
FOOD-CT-2006-016236: Network of Excellence for Epizootic Disease Diagnosis and Control. Acronym:
EPIZONE (2006-2009).
Web address: http://www.epizone-eu.net/
Role: Partner
SSP5-044429: Improved Diagnosis and Early Warning Systems for Avian Influenza Outbreak Management.
Acronym: FLUTEST (2007-2010).
Role: Partner
SSPE-044212: International training course in molecular diagnostics of influenza virus directed towards INCO
countries. Acronym FLUTRAIN (2007-2010)
Web address: http://www.flutrain.eu
Role: Partner
SSP5-044266: The EU, China and South East Asia Dialog for the Development of Research Areas in Animal
Health of Mutual Interest. Acronym: ASEMDialog (2007-2008)
Role: Partner
SSP5-04462: Capacity building for the control of Avian influenza through technology transfer and training
directed towards INCO countries. Acronym: ConFluTech (2007-2010)
Role: Partner
In these projects a high number of European partner laboratories, external partners and subcontractors work
together on various subjects, including the international standardisation and harmonisation of PCR-based
diagnostic assays.
The total number of partners participating in the listed projects:
MULTIPLEX PCR:
LAB-ON-SITE:
CSFVACCINE & WILDBOAR:
MED-VET-NET:
PATHOGEN COMBAT:
EPIZONE
FLUTEST
FLUTRAIN
ConFluTech
AsemDialog
6 partners
9 partners
9 partners
15 partners
42 partners
20 partners
17 partners
12 partners
15 partners
13 partners
Detailed information on international standardisation and validation is indicated on the web sites of the projects,
including material and SOP transfer, ring tests, by following the five stages of OIE validation.
2.
Proposal or development of any procedure that will facilitate harmonisation of international
regulations applicable to the surveillance and control of animal diseases, food safety and animal
welfare
Authors of a new chapter (1.1.4) on the Validation and quality control of polymerase chain reaction methods used
for the diagnosis of infectious diseases, to be published in the OIE Manual of Diagnostic Tests and Vaccines for
Terrestrial Animals, Sixth Edition, 2008.
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Biotechnology-Based Diagnosis of Infectious Diseases in Veterinary Medicine
Authors of a new chapter on Validation and quality control of polymerase chain reaction methods used for the
diagnosis of infectious diseases, to be included in the new edition of the booklet "OIE Quality Standard and
Guidelines for Veterinary Laboratories: Infectious Diseases"
3.
Placement of expert consultants at the disposal of the Office International des Epizooties
Consultant expertise was provided to OIE, IAEA and FAO by regular change of information and by participation
in meetings, see below.
Chairmanship in the work of the OIE ad hoc Group on Biotechnology, Paris, France, regular meetings, preparation
of recommendations and reports in biotechnology issues.
Participation in the work of the European Platform for Global Animal Health, Brussels, Belgium, regular
meetings, preparation of recommendations and reports in issues of Transboundary Animal Diseases.
Participation in the work of the Animal Production and Health Section, FAO/IAEA Joint Division, IAEA, Vienna
(sister Collaborating Centre, see above). Development of novel diagnostic methods, international standardisation
and validation, providing SOPs to member countries, international training.
Areas of expertise
 Research, novel biotechnology-based techniques (e.g., microarrays, isothermal amplification, rapid
sequencing) aimed for diagnostic use
 Diagnostic real-time PCR (e.g., TaqMan, Molecular beacons, Primer-Probe Energy Transfer methods)
 Laboratory automation, molecular diagnostics
 Quality assurance and validation of molecular assays
Other activities related to the mandate
of OIE Collaborating Centres
4.
Provision of scientific and technical training, within the remit of the mandate given by the OIE, to
personnel from Member Countries of the Office
The laboratory provided training for scientists visiting from various countries to Uppsala. In addition, the
laboratory housed prominent experts, in order to exchange ideas and improve the level of molecular diagnosis.
1)
Dr. Ákos Hornyák, Central Veterinary Institute, Budapest, Hungary
Visit: four months. Subject: exchange of ideas in PCR based molecular diagnosis, development of a novel
real-time PCR assay for the improved detection of coronaviruses and other pathogens of zoonotic
importance.
2)
Dr. Tomasz Stadejek, National Veterinary Research Institute, Pulawy, Poland
Visit: two weeks. Subject: exchange of ideas in PCR based molecular diagnosis, focusing on improved
detection of CSFV and PRRSV.
3)
Dr. Sandra Cuevas Romero, Centro Nacional de Investigaciones Disciplinarias en Microbiologia Animal
INIFAP. México D.F, Mexico
Visit: two months. Subject: training and development of a novel real-time PCR assay for the improved
detection of porcine rubula virus (LPMV).
4)
Dr. Wai Kwan Wong, Virology Branch, Agri-Food & Veterinary Authority, Singapore
Visit: two weeks. Subject: training and discussion regarding diagnosis of HPAI with emphasis on BSL-3
practices.
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Biotechnology-Based Diagnosis of Infectious Diseases in Veterinary Medicine
5.
5)
Dr. Adám Bálint, Central Veterinary Institute, Budapest, Hungary
Visit: Four months. Subject: exchange of ideas in PCR based molecular diagnosis, coronaviruses and other
pathogens of zoonotic importance, including highly influenza viruses.
6)
Dr. Attila Farsang, Institute for Veterinary Medicinal Products, Budapest, Hungary
Visit: Three months. Subject: exchange of ideas in PCR based molecular diagnosis, coronaviruses and other
pathogens of zoonotic importance.
Organisation of scientific meetings on behalf of the Office
None
6.
Coordination of scientific and technical studies in collaboration with other laboratories or
organisations
The Collaborating Centre is co-ordinator or partner in nine ongoing EU projects (see above). In the frame of these
projects large international networks collaborate on development of new molecular diagnostic methods,
international standardisation and validation (see web sites of the projects for details).
7.
Publication and dissemination of any information within the remit of the mandate given by the
OIE that may be useful to Member Countries of the Office
Scientific publications
Antal M, Farkas T, Germán P, Belák S, Kiss I. Real-time reverse transcription-polymerase chain reaction detection
of Newcastle disease virus using light upon extension fluorogenic primers. J Vet Diagn Invest. 2007 Jul;
19(4):400-4.
Banér J, Gyarmati P, Yacoub A, Hakhverdyan M, Stenberg J, Ericsson O, Nilsson M, Landegren U, Belák S.
Microarray-based molecular detection of foot-and-mouth disease, vesicular stomatitis and swine vesicular disease
viruses, using padlock probes. J Virol Methods. 2007 Aug; 143(2):200-6.
Belák S. Experiences of an OIE Collaborating Centre in molecular diagnosis of transboundary animal diseases: a
review. Dev Biol (Basel). 2007; 128: 103-12.
Belák S. Molecular diagnosis of viral diseases, present trends and future aspects A view from the OIE
Collaborating Centre for the Application of Polymerase Chain Reaction Methods for Diagnosis of Viral Diseases
in Veterinary Medicine. Vaccine. 2007 Jul 26; 25(30):5444-52. Epub 2006 Dec 28. Review.
Blomström AL, Hakhverdyan M, Reid SM, Dukes JP, King DP, Belák S, Berg M. A one-step reverse transcriptase
loop-mediated isothermal amplification assay for simple and rapid detection of swine vesicular disease virus. J
Virol Methods. 2008 Jan; 147(1):188-193.
Escutenaire S, Mohamed N, Isaksson M, Thorén P, Klingeborn B, Belák S, Berg M, Blomberg J. SYBR Green
real-time reverse transcription-polymerase chain reaction assay for the generic detection of coronaviruses. Arch
Virol. 2007 Jan; 152(1):41-58.
Farkas T, Antal M, Sámi L, Germán P, Kecskeméti S, Kardos G, Belák S, Kiss I. Rapid and simultaneous
detection of avian influenza and newcastle disease viruses by duplex polymerase chain reaction assay. Zoonoses
Public Health. 2007; 54(1):38-43.
Fernández J, Agüero M, Romero L, Sánchez C, Belák S, Arias M, Sánchez-Vizcaíno JM. Rapid and differential
diagnosis of foot-and-mouth disease, swine vesicular disease, and vesicular stomatitis by a new multiplex RT-PCR
assay. J Virol Methods. 2008 Feb; 147(2): 301-11.
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Biotechnology-Based Diagnosis of Infectious Diseases in Veterinary Medicine
Gyarmati P, Mohammed N, Norder H, Blomberg J, Belák S, Widén F. Universal detection of hepatitis E virus by
two real-time PCR assays: TaqMan and Primer-Probe Energy Transfer. J Virol Methods. 2007 Dec; 146(1-2):22635.
Kiss I, Germán P, Sámi L, Antal M, Farkas T, Kardos G, Kecskeméti S, Dán A, Belák S. Application of real-time
RT-PCR utilising lux (light upon extension) fluorogenic primer for the rapid detection of avian influenza viruses.
Acta Vet Hung. 2006 Dec; 54(4): 525-33.
Liu L, Widén F, Baule C, Belák S. A one-step, gel-based RT-PCR assay with comparable performance to realtime RT-PCR for detection of classical swine fever virus. J Virol Methods. 2007 Feb; 139(2): 203-7.
McKillen J, Hjertner B, Millar A, McNeilly F, Belák S, Adair B, Allan G. Molecular beacon real-time PCR
detection of swine viruses. J Virol Methods. 2007 Mar; 140(1-2):155-65.
Sámi L, Ursu K, McKillen J, Kecskeméti S, Belák S, Kiss I. Simultaneous detection of three porcine viruses by
multiplex PCR. Acta Vet Hung. 2007 Jun; 55(2):267-76.
Wensman JJ, Thorén P, Hakhverdyan M, Belák S, Berg M. Development of a real-time RT-PCR assay for
improved detection of Borna disease virus. J Virol Methods. 2007 Jul; 143(1): 1-10.
Presentations at international conferences and meetings
Belák, S.: Early and rapid diagnosis of transboundary animal diseases (TADs), such as avian influenza. A view
from the “OIE Collaborating Centre for the application biotechnology-based methods for diagnosis of infectious
diseases in veterinary medicine”. First RCM, IAEA, Vienna 19-23 March, 2007. Oral presentation.
LeBlanc, N., Gantelius, J., Schwenk, J.M, Ståhl, K., Andersson-Svahn, H., Belák, S.: Viral Detection using
Suspension and Magnetic Bead Microarrays. Swedish Molecular Virology Network, Annual Meeting, Skytteholm,
Sweden 7-8 May, 2007. Oral presentation.
Sandwall, E., Wensman, J.J., Blomström, A.-L., Riberdahl, E., Johannisson, A., Belák, S., Berg, A.-L. & Berg, M.
2007. Borna disease virus infection inhibits apoptosis in persistently infected rat astrocytoma cells. 2nd Meeting of
the Swedish Molecular Virology Network, Skytteholm, Sweden, 7-8 May 2007. Oral presentation.
Blomström, A.-L., Hakhverdyan, M., Belák, S. & Berg, M. 2007. Development of a RT-LAMP assay for the
detection of swine vesicular disease virus. 2nd Meeting of the Swedish Molecular Virology Network, Skytteholm,
Sweden, 7-8 May 2007. Poster presentation.
Blomström, A.-L., Hakhverdyan, M., Belák, S. & Berg, M. 2007. Development of a RT-LAMP assay for the
detection of swine vesicular disease virus. First annual meeting Epizone "Epizone inside out", Lublin, Poland, 30th
May- 1st June 2007. Poster presentation.
Banér, J., Gyarmati, P., Yacoub, A., Hakhverdyan, M., Stenberg, J., Ericsson, O., Nilsson, M., Landegren, U.,
Belák, S.: Utilization of a microarray-based padlock probe system in the viral diagnostics. First annual meeting
Epizone "Epizone inside out", Lublin, Poland, 30th May- 1st June 2007. Poster presentation.
Hakhverdyan M., Rasmussen T. B, Uttenthal Å., Belák S.: One-step primer-probe energy transfer (PriProET) realtime RT-PCR detection of SVDV using Rotor-Gene 6000. Proceedings of the 13th International World
Association of Veterinary Laboratory Diagnosticians Symposium (WAVLD), 11-14 November 2007, Melbourne,
Australia, p. 174.
LeBlanc, N., Gyarmati, P., Belák, S.: Detection and subtyping of avian influenza on a suspension microarray.
Epizone WP 4.2 DNA chip meeting, Copenhagen 11 December, 2007. Oral presentation.
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