OIE Reference Laboratory Reports
Activities in 2010
Name of disease (or topic) for
which you are a designated OIE
Reference Laboratory:
Address of laboratory:
Enzootic bovine leukosis
Faculty of Veterinary Medicine
University of Leipzig
An den Tierkliniken 29; 04103 Leipzig/Germany
Tel.:
+49(341) 973.82.00
Fax:
+49(341) 973.82.19
e-mail address:
thomas.vahlenkamp@uni-leipzig.de
website:
www.vetmed.uni-leipzig.de
Name of Head of Laboratory
(Responsible Official):
Name of OIE Reference Expert:
Prof. Dr. Dr. Thomas W. Vahlenkamp
Name of writer of this report
(if different from above):
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
1
Enzootic bovine leukosis
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
The OIE and national reference laboratory for Enzootic Bovine Leukosis (EBL) at the Friedrich-Loeffler-Institute
uses different commercial test kits for the detection of anti-Bovine Leukemia Virus (BLV) antibodies. The test
systems for serum are enzyme linked-immunosorbent assays (ELISA) and Agargel-Immunodiffusion (AGID)
tests. Test systems for the detection of antibodies in milk samples are ELISA. In addition to a nested PCR for the
detection of provirus in tissue samples of serologically doubtful animals a real time PCR has been established for
the detection of proviral DNA in PBMC samples of infected cattle.
During the reporting period seven batches from different ELISAs and one batch from an AGID were evaluated for
their national use in Germany.
The number of samples sent to our laboratory in 2010 and verified for the presence of anti-BLV specific
antibodies and/or proviral DNA is given in the table below.
2.
Test
For
Specificity
Total
ELISA (confirmation test)
Antibody detection
BLV gp51, p24
137
Agargel-Immunodiffusion (AGID) test
Antibody detection
BLV antigen
10
Polymerase chain reaction (PCR)
Provirus detection
Viral envelope gene
29
Real time
polymerase chain reaction (PCR)
Provirus detection
Viral polymerase gene
25
Production and distribution of diagnostic reagents
The new international reference was distributed to the OIE reference laboratories at the Veterinary Laboratories
Agency (VLA) in Weybridge/UK and at the National Veterinary Research Institute (NVRI) in Pulawy/Poland.
The new reference serum E-05 which was produced at the Friedrich-Loeffler-Institut was distributed upon request
to national diagnostic laboratories in 10 countries including Latvia, Morocco, Ireland, Germany (2x), Poland,
Switzerland, Cyprus, Czech republic, Spain, Estonia.
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
In the reporting period the use of the new generated real time PCR for the detection of BLV genome was
introduced into the OIE Manual of Diagnostic Tests and Vaccines for Terrestrial Animals (Terrestrial Manual)
Chapter 2.4.11, Enzootic Bovine Leukosis.
4.
Preparation and supply of international reference standards for diagnostic tests or vaccines
In a continuing effort the EBL reference serum E-05 was prepared in aliquots of 1 and 2 ml and retested for the
presence of anti-BLV antibodies at the Friedrich-Loeffler-Institut.
2
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
Enzootic bovine leukosis
In the reporting period we distributed the reference serum E-05 upon request to national diagnostic laboratories in
10 countries including Latvia, Morocco, Ireland, Germany (2x), Poland, Switzerland, Cyprus, Czech Republic,
Spain, Estonia.
5.
Research and development of new procedures for diagnosis and control
The validation of a new real time RT-PCR method for the detection BLV in peripheral blood of infected cattle was
finalised during the reporting period. The assay was validated with 350 samples of bovine peripheral blood
mononuclear cells including 144 samples from BLV-seropositive animals worldwide (South America, Europe,
Middle East, Australia) representing 5 out of the 7 BLV genotypes recently described. The validation showed that
the new real time RT-PCR based on the BLV-polymerase gene detects samples from different clusters and is
therefore suited to be introduced as reference method in the OIE diagnostic manual. The publication of the PCR is
submitted.
6.
Collection, analysis and dissemination of epizootiological data relevant to international disease
control
During the reporting period there were no significant new information with regard to the epizootiological situation
of Enzootic Bovine Leukosis.
7.
Provision of consultant expertise to OIE or to OIE Members
No activities.
8.
Provision of scientific and technical training to personnel from other OIE Members
No activities.
9.
Provision of diagnostic testing facilities to other OIE Members
No activities.
10. Organisation of international scientific meetings on behalf of OIE or other international bodies
A meeting of the OIE-reference laboratories for EBL was held at the Friedrich-Loeffler-Institut on February 25th
and 26th 2010. Presentations of all OIE reference laboratories were discussed. It was agreed to introduce the real
time PCR established at the Friedrich-Loeffler-Institute as a possible method for the detection of proviral DNA in
the OIE Manual of Diagnostic Tests and Vaccines for Terrestrial Animals (Terrestrial Manual) Chapter 2.4.11,
Enzootic Bovine Leukosis.
11. Participation in international scientific collaborative studies
We included in the validation study for the real time RT-PCR method samples derived from our collegues from
the other OIE-reference laboratories. We also participated in a serological ring trial organised by the OIE reference
laboratory in Poland with the ELISA test kit from IDEXX and the AGID from Synbiotics.
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

Vahlenkamp, T.W.: Real time PCR for the detection of EBL. OIE Reference Laboratory Meeting at the
Friedrich-Loeffler-Institut, Greifswald, Insel Riems on February 25th and 26th 2010.
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
3
Enzootic bovine leukosis

Vahlenkamp, T.W.: Aktuelle Tierseuchensituation in Deutschland; Globalisierung und Klimawandel - Welche
Infektionsrisiken ergeben sich? Deutsche Veterinärmedizinische Gesellschaft (DVG), Weiterbildungsmodul
‘Infektionskrankheiten’, Klinik für Klauentiere, Freie Universität Berlin on January 30 th 2010.

Scientific publications in peer-reviewed journals
Manuscript submission (Heenemann et al.: Development of a real-time polymerase chain reaction for detecting
bovine leukemia virus [BLV]).

Other communications
Give details
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