OIE Reference Laboratory Reports
Activities in 2011
Name of disease (or topic) for
which you are a designated OIE
Reference Laboratory:
Address of laboratory
Enzootic bovine leukosis
National Veterinary Research Institute
Al. Partyzantow 57
24-100 Pulawy,
POLAND
Tel.:
+48-818893114
Fax:
+48-818862595
e-mail address:
jkuzmak@piwet.pulawy.pl
www.piwet.pulawy.pl
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
Dr Krzysztof Niemczuk,
Director general of NVRI
Jacek Kuźmak, DVM, PhD, Professor
The same as above
1
Enzootic bovine leukosis
Part I: Summary of general activities related to the disease
1.
2.
Test(s) in use/or available for the specified disease/topic at your laboratory
Test
For
Specificity
Total
ELISA screening
Antibody
Group
1399
ELISA verification
Antibody
Group
204
AGID
Antibody
Group
518
Nested PCR/real time PCR
Proviral DNA
Group
16
Production and distribution of diagnostic reagents
Amount supplied nationally
(including for own use)
Amount supplied to
other countries
P9 Control positive serum
(substandard to E05 OIE standard)
160 ml
15 ml
N27 control negative serum
366 ml
13 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
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
The laboratory participated quarterly in the proficiency tests organized by Quality Control Unit, Sutton
Bonington, VLA (UK) and the results all tests were satisfactory.
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
The laboratory organized one round of proficiency test for 28 regional laboratories in Poland.
10 lyophilizated bovine sera, including specimens from three uninfected and seven BLV-infected cattle were
sent to these labs and were tested by both AGID and ELISA. The Se and Sp of this test was 100%.
The same sera were sent to 19 laboratories from 13 countries, including national reference laboratories and
laboratories from R&D of some commercial companies producing diagnostic kits. The results were identical
with expected ones for all labs working with ELISA and discrepant for two labs, out of 15, working with
AGID.
4.
Preparation and supply of international reference standards for diagnostic tests or vaccines
N/A
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Annual reports of OIE Reference Centres, 2011
Enzootic bovine leukosis
5.
Research and development of new procedures for diagnosis and control
Laboratory has participated in the evaluation of sensitivity and specificity of new developed real-time PCR
method for detection of proviral DNA of BLV, regarding further application of this test for diagnosis of BLV
infection. Peripheral blood leukocytes were prepared from 15 cattle, seropositive for BLV, from different region
from Ukraine and were tested under collaboration with diagnostic laboratory in Kharkov.
Under collaboration with VLA AH in Weybridge the laboratory developed a new real time PCR based on
detection of pol gene sequence, using TaqMan probe.
6.
Collection, analysis and dissemination of epizootiological data relevant to international disease
control
N/A
7.
Maintenance of a system of quality assurance, biosafety and biosecurity relevant to the
pathogen and the disease concerned
The NVRI has implemented quality control system according to the ISO norm 17025. Regarding EBL all
diagnostic procedures i.e. AGID and ELISA are accredited since 2009 by Polish Center for Accreditation (PCA) –
accreditation no AB1090. The laboratory is located in a part of facility with CL2 biosafety level.
8.
Provision of consultant expertise to OIE or to OIE Member Countries
The laboratory participated in the revision of updated version of Chapter on Enzootic Bovine Leukosis in OIE
Terrestrial Manual
9.
Provision of scientific and technical training to personnel from other OIE Member Countries
Dr Natalya Kozyrieva from All-Russia Y.Kovalenko Institute of Experimental Veterinary Medicine, Moscov,
Russia was trained at the NVRI on October 4-14, 2011 regarding application of serological (ELISA and AGID)
and molecular methods for detection of BLV infection (nested PCR, real time PCR, RFLP, isolation and
purification of PBL and PBMC from peripheral blood) and EU legislation in terms of official control of EBL.
10. Provision of diagnostic testing facilities to other OIE Member Countries
N/A
11. Organisation of international scientific meetings on behalf of OIE or other international bodies
The laboratory stayed in touch with other OIE refrerence laboratory (VLA) and exchanged the data on
organization of official meeting of OIE reference laboratories in 2012 .
12. Participation in international scientific collaborative studies
The laboratory in collaboration with: Urals State Scientific Research Institute of Veterinary Medicine,
Ekaterinburg, Russia , All Russian Research Institute of Animal Breeding, Moscow, Russia, National Belarusian
Veterinary Center, Minsk, Belarus, National Scientific Center Institute of Experimental and Clinical Veterinary
Medicine, Kharkov, Ukraine developed a project on genetic characterization of BLV strains isolated from cattle
from Eurasia. The 444 bp fragment of env gene were amplified by nested PCR from 44 isolates of BLV from
Ukraine (9 isolates), Russia (21), Belarus (2) and Poland (12). The respective fragments were subjected to
sequencing and phylogenetic analysis using Neighbour Joining and Bayes methods. The results showed that the
Annual reports of OIE Reference Centres, 2011
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Enzootic bovine leukosis
sequences were mainly located within genotype 4 and 7, however, some sequences created a new genotype 8. It
seems that this genotype is typical for BLV isolates circulated in cattle from South-East Europe.
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
J. Kuzmak, lecture on “Molecular analysis of BLV isolates come from Russia and other countries – a link to an
epidemiological study” was presented on International Seminar at Urals State Scientific Research Institute of
Veterinary Medicine, Ekaterinburg, Russia, 14-18 June, 2011
Rola M., Kozaczyńska B., Pluta A., Kuźmak J. Bovine Immunodeficiency Virus as a potentiating cofactor for the
experimental Bovine Leukemia Virus infection in sheep. 15th International Conference on Human Retrovirology
HTLV and Related Viruses, 4-8th June, 2011, Leuven/Gembloux, Belgium, poster presentation
Szczotka M., Kuźmak J., Telomerase activity in cattle infected with bovine leukemia virus (BLV), 29th Meeting
of the European Society of Veterinary Pathology and European College of Veterinary Pathologists (ESVP/ECVP),
6 – 11.09.2011, Uppsala, Sweden, poster presentation
Szczotka M., Kuźmak J., Determination of telomerase activity in cattle infected with bovine leukemia virus. 23rd
Workshop on Retroviral Pathogenesis: 2-5.11.2011, Montpellier, France, oral presentation

Scientific publications in peer-reviewed journals
Szczotka M., Kuźmak J., Kostro K., Iwan E., Telomerase activity in cattle infected with bovine leukaemia virus.
Bull Vet Inst Pulawy, 2011, 55, No 4, 587-592
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Annual reports of OIE Reference Centres, 2011