Real-time PCR

advertisement
OIE Reference Laboratory Reports
Activities in 2011
Name of disease (or topic) for
which you are a designated OIE
Reference Laboratory:
Address of laboratory:
Capripoxvirus Reference Laboratory
Institute for Animal Health
Ash Road, Pirbright
Surrey, GU24 0NF
UNITED KINGDOM
Tel.:
+44 (0)1483 232441
Fax:
+44 (0)1483 232448
e-mail address:
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):
Annual reports of OIE Reference Centres, 2011
Lumpy skin disease
iah@iah.ac.uk, eeva.tuppurainen@iah.ac.uk
http://www.iah.bbsrc.ac.uk
Dr Michael Johnson
Eeva Tuppurainen,
Head of Capripoxvirus
Reference Laboratory,
Institute for Animal Health, Pirbright
Same as above
1
Lumpy skin disease
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
Electron microscopy
Antigen
Group
0
SNT
Antibody
Group
178
VN
Antibody
Group
Virus isolation on lamb
testis and bovine dermis
primary cell culture
Antigen
Gel-based PCR
Real-time PCR
2.
102
Antigen
Group
Antigen
Group
425
Production and distribution of diagnostic reagents
Amount supplied nationally
(including for own use)
Amount supplied to other
countries
Control positive serum
No pos antiserum production required
during reporting period
35 ml
Live virus
5 ml
1 x 1 ml in freeze dried form
Extracted DNA from LSD virus
(different strains)
2 ml
0 ml
Type of reagent
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
The following institutes were provided with method sheets for the diagnostic tests which are currently used at the
Capripoxvirus Reference Laboratory at IAH for the detection capripoxvirus antigen or antibody: Animal Health
Development Programme, Ministry of Agriculture, Irrigation and Livestock, CVDRL, Dar-ul-Aman, Kabul,
Afganistan
A previously published real-time PCR (Bowden et al., 2008) and a robotic extraction method were adapted to a highthroughput system and validated (Stage 2). The results were published in the Journal of Virological Methods (DOI
10.1016/j.jviromet.2011.11.015)
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
A ring trial on the performance of serum neutralisation test was successfully carried out by the reference
laboratories for LSD at the Institute for Animal Health, UK and the Onderstepoort Veterinary Institute (OVI),
2
Annual reports of OIE Reference Centres, 2011
Lumpy skin disease
SA. We are continuing to strengthen our contact and improve harmonisation in testing between our reference
laboratories.
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
No activity during the reporting period
4.
Preparation and supply of international reference standards for diagnostic tests or vaccines
No international standard reagents are available for capripoxviruses.
The following institutes were provided with LSDV-positive antiserum raised in experimentally infected cattle at
the IAH, to be used as a positive control in serological tests for the detection of capripoxvirus antibodies: CODACERVA, Department of General Virology, Section of Epizootic Diseases, Belgium (5 ml) and Laboratoire de
Production des Vaccins Antirabique et de la Clavelee, Institut Pasteur Algerie, 1, Rue du Caire Kouba,
16032 Algerie (5 ml).
CSIRO Livestock Industries, Australia Animal Health Laboratory (AAHL), was provided with 410 x 50 µl
capripoxvirus positive antiserum, comprising field samples and serum collected from experimentally infected and
vaccinated animals (cattle, sheep and goats). These samples will be used for the development of a new ELISA for
capripoxviruses.
OIE reference laboratory for LSD at the Onderstepoort Veterinary Institute (OIE), South Africa, was provided
with LSD antiserum (a panel of 10 samples) to evaluate the serum neutralisation test for the quality control
purposes and for the inter-laboratory test evaluation.
The following institutes were provided with LSDV (Neethling strain) for the infection of experimental cattle and
/or for the development/set up of diagnostic assays for LSD antigen or antibodies: CODA-CERVA, Department of
General Virology, Section of Epizootic Diseases, Belgium (1 ml in freeze dried form).
5.
Research and development of new procedures for diagnosis and control
A previously published real-time PCR (Bowden et al., 2008) and a robotic extraction method were adapted to a highthroughput system and validated (Stage 2). The results were published in the Journal of Virological Methods (DOI
10.1016/j.jviromet.2011.11.015)
The performance of a LAMP assay, recently developed at IAH for capripoxviruses, was evaluated using field and
experimental samples and different capripox virus isolates. These isolates were previously tested at the capripox
reference laboratory using both conventional and real-time PCR assays.
A collaboration project entitled «Evaluation of a recombinant antigen ELISA for detection of antibodies against
capripoxviruses» by the Capripoxvirus Reference Laboratory, IAH, the Roslin Institute, the University of Edinburg,
CSIRO Livestock Industries (CLI), Australian Animal Health Laboratory (AAHL) and Canadian Food Inspection
Agency (CFIA) National Centre for Foreign Animal Disease (NCFAD) is on-going. A number of new antigens have
been identified based on the capripoxvirus protein microarray data at CSIRO. Serum samples (410), comprising field
samples and samples from vaccinated and experimentally infected animals (cattle, sheep and goats), held at the
Capripoxvirus Reference Laboratory stocks, were sent to CSIRO, for the evaluation of new antigen candidates.
6.
Collection, analysis and dissemination of epizootiological data relevant to international disease
control
Epidemiological surveys: A project entitled “Transmission of lumpy skin disease virus by ixodid tick-vector” is
on-going in collaboration with the Department of Veterinary Tropical Diseases, Faculty of Veterinary Science,
University of Pretoria. Transmission of the LSDV by tick vectors was demonstrated and the first results of the
Annual reports of OIE Reference Centres, 2011
3
Lumpy skin disease
study were presented at the 30th World Veterinary Congress in Cape Town, SA, in October 2011. A publication is
in progress.
7.
Maintenance of a system of quality assurance, biosafety and biosecurity relevant to the
pathogen and the disease concerned
Capripoxvirus reference laboratory is accredited to ISO 9001, real-time PCR assay for capripoxviruses to
ISO17025.
8.
Provision of consultant expertise to OIE or to OIE Member Countries
A twinning project is on-going with the National Animal Disease Diagnostics and Epidemiology Centre
(NADDEC), Ministry of Agriculture Animal Industry and Fisheries, Entebbe, Uganda. Drs E. Tuppurainen and M.
Henstock from IAH capripoxvirus reference laboratory, visited Uganda 20-25th of June 2011. Conventional and
real-time PCR methods for capripoxviruses were set up at NADDEC laboratory. The performance of these tests
will be followed-up by testing of a panel of duplicate field samples in parent and candidate laboratories in early
2012.
Upon request OIE member countries were advised on the availability of tests for the detection of capripoxvirus
antigen or antibodies or provided with method sheets for setting up diagnostic methods.
Lumpy skin disease data sheets were reviewed for CABI Animal Health and Production Compendium.
9.
Provision of scientific and technical training to personnel from other OIE Member Countries
The following institutes/countries sent participants to attend a training course at IAH Pirbright Laboratory, UK
(3.05-13.05.11) entitled “Bluetongue, Peste des Petits Ruminants (PPR), and Capripox Diagnostics Course“:
National Animal Disease Diagnostics and Epidemiology Centre (NADDEC), Entebbe, Uganda (2) and All-Russia
Scientific Institute of Veterinary Virology and Microbiology, Pokrov, Russia (1).
A twinning project with the National Animal Disease Diagnostics and Epidemiology Centre (NADDEC), Ministry
of Agriculture Animal Industry and Fisheries, Entebbe, Uganda commenced on 1.11.2010. The project includes
capripoxviruses with the aim to train the personnel and upgrade the diagnostic facilities in the Ugandan lab.
Capripoxvirus presentation was included into the Notifiable and Exotic Disease Course which was organized for
UK government veterinarians (30.06-1.07.11) by IAH.
10. Provision of diagnostic testing facilities to other OIE Member Countries
NADDEC Uganda sent 39 tissue and serum samples for capripoxviruses for confirmatory diagnostic testing
11. Organisation of international scientific meetings on behalf of OIE or other international bodies
No activity during this reporting period
12. Participation in international scientific collaborative studies
Collaboration with Koret School of Veterinary Medicine, Hebrew University of Jerusalem, Israel on the
transmission of LSDV by biting flies and mathematical modelling of LSD was completed and the results were
published.
Collaboration with the University of Liverpool, UK, entitled « Comparison of the whole genome of Egyptian old
(1993) LSD isolate with Egyptian new (2006) and Israel 2007 LSD strains is on-going.
4
Annual reports of OIE Reference Centres, 2011
Lumpy skin disease
A 3-year project entitled “Transmission of lumpy skin disease virus by ixodid tick-vector” is on-going in
collaboration with the Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of
Pretoria.
A research project on the evaluation of the efficacy of 3 capripoxvirus live vaccine strains against LSD in Ethiopia
has been initiated. The project is a collaboration between the National Animal Health Diagnostic and Investigation
Center (NAHDIC), Ethiopia, National Veterinary Institute (NVI), Ethiopia, Centre for International Research for
Agricultural Development (CIRAD), France, and Capripoxvirus Reference Laboratory, Institute for Animal
Health, UK.
A collaboration project entitled «Evaluation of a recombinant antigen ELISA for detection of antibodies against
capripoxviruses» by the Capripoxvirus Reference Laboratory, IAH, the Roslin Institute, University of Edinburg,
CSIRO Livestock Industries (CLI), Australian Animal Health Laboratory (AAHL) and Canadian Food Inspection
Agency (CFIA) National Centre for Foreign Animal Disease (NCFAD) is on-going. A number of new potential
antigens for ELISA have been identified based on the capripoxvirus protein microarray data at CSIRO.
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
Oral presentation was given by Dr E. Tuppurainen, entitled “Determination of the role of ixodid (hard) ticks in the
transmission of lumpy skin disease virus” at the 30th World Veterinary Congress, 10-14th Oct 2011, Cape Town,
South Africa

Scientific publications in peer-reviewed journals
Lamien C. E., Le Goff C., Silber R., Wallace D. B., Gulyaz V., Tuppurainen E., Madani H., Caufour P., Adam
T., Harrak M. E., Luckins A. G., Albina E., Diallo A. (2011). Use of the Capripoxvirus homologue of Vaccinia
virus 30kDa RNA polymerase subunit (RPO30) gene as a novel diagnostic and genotyping target: Development of
a classical PCR method to differentiate Goat poxvirus from Sheep poxvirus. Veterinary Microbiology, 149 (1-2),
30-39.
Lamien C.E., Lelenta M., Goger W., Silber R., Tuppurainen E., Matijevic M., Luckins A.G. and Diallo A.
(2011). Real time PCR method for simultaneous detection, quantitation and differentiation of Capripoxviruses,
Journal of Virological Methods, 171 (1), 134-40.
E.S.M. Tuppurainen and C.A.L. Oura (2011) Lumpy skin disease: an emerging threat to Europe, the
Middle East and Asia. Transboundary and Emerging Diseases., in press, DOI: 10.1111/j.18651682.2011.01242.x
Stubbs S., Oura C.A.L., Henstock M., Bowden T.R., King D.P. and Tuppurainen E.S.M. (2011). Validation of a
high-throughput real-time polymerase chain reaction assay for the detection of capripoxviral DNA. Journal of
Virological Methods, in press, DOI 10.1016/j.jviromet.2011.11.015
Magori-Cohen R., Louzoun Y., Hertziger Y., Oron E., Arazi A., Tuppurainen E., Shpigel N.Y. and Klement E.
(2011) Mathematical modelling and evaluation of the different routes of transmission of lumpy skin disease virus.
Veterinary Research, accepted, DOI: 10.1186/1297-9716-43-1

Other communications
Lumpy skin disease data sheets were reviewed for CABI Animal Health and Production Compendium
_______________
Annual reports of OIE Reference Centres, 2011
5
Download