Enzootic bovine leukosis
Activities in 2009
Enzootic bovine leukosis
Jacek Kuzmak
National Veterinary Research Institute
24-100 Pulawy, Poland
Tel.: (+48-81) 889 3114, Fax: (+48-81) 886 2595
, www.piwet.pulawy.pl
Summary of general activities related to the disease
1. Test(s) in use/or available for the specified disease at your laboratory
Test / For / Specificity / TotalELISA screening / Antibody / Group / 2320
ELISA verification / Antibody / Group / 127
AGID / Antibody / Group / 29
Nested PCR
/ Proviral DNA / Group / 142. Production and distribution of diagnostic reagents
N27 reference serum which is national standard of negative serum for AGID test for enzootic bovine leucosis and P9 reference serum which is substandard to E05 reference positive serum for ELISA and AGID
No production of these sera took place in 2009; however, current stock includes about 2000 vials of N27 and 1500 vials of P9 serum.
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
Laboratory has participated in the evaluation of real-time PCR method for detection of proviral DNA of BLV, with regard to further development for diagnosis of BLV infection. DNA samples and peripheral blood leukocyte preparations from 76 field animals have been tested in reference laboratory at VLA.
4. Preparation and supply of international reference standards for diagnostic tests or vaccines
N/A
5. Research and development of new procedures for diagnosis and control
A new format of real-tme PCR was validated for detection of proviral DNA of BLV. Primers selected from pol gene region of BLV and TaqMan probe labeleled with 6-FAM and BHQ1 used as a quencher were designed to test DNA samples from 14 cattle with a strong antibody response again BLV by ELISA and 57 animals which had given rise to borderline and unexpected ELISA results. The real-time assay showed a detection limit of one copy of DNA-BLV and in comparison to the conventional PCR detected approximately 5% more specimens positive. This test help elucidate the disease status of animals when borderline ELISA results are obtained in the diagnostic laboratory.
6. Collection, analysis and dissemination of epizootiological data relevant to international disease control
N/A
7. Provision of consultant expertise to OIE or to OIE Members
N/A
8. Provision of scientific and technical training to personnel from other OIE Members
Dr Oksana Blotskaya a scientist from the Institute of Clinical and Experimental Veterinary Medicine in Kharkov, Ukraine was trained in the reference laboratory at NVRI on May 4-9, 2009. Training concerned preparation and validation of a new Ukrainian national standard of positive BLV serum. Three scientist from Urals State Scientific Research Institute of Veterinary Medicine, Ekaterinburg, Rosja (Ewgienia Szilowa, Sofia Satchikowa, Maksim Petropawlosky) were trained at NVRI on June 1-18, 2009 regarding application different ELISA tests for serological diagnosis of BLV, batch control of AGID and ELISA tests available on the market, EU legislation in terms of official control of EBL. Dr Davor Baliĉ from Croation Veterinary Institute , Department in Vinkovci, Croatia was trained on October 12-16, 2009 in terms of application different ELISA tests for serological diagnosis of BLV and batch control of AGID and ELISA tests available on the market.
9. Provision of diagnostic testing facilities to other OIE Members
N/A
10. Organisation of international scientific meetings on behalf of OIE or other international bodies
No official meeting of OIE reference laboratories was organized in 2009, however, after discussion with Dr T.Vahlencamp, FLI next meeting will be organized by FLI, Insel Riems on February 2010.
11. Participation in international scientific collaborative studies
The project No WAR/342/121 “ Improvement of diagnosis of infection with bovine retroviruses: BLV, BFV and BIV” supported by British Council was developed in collaboration with Dept. Virology,VLA .Weybridge. A set of of primers selected from pol gene region of BLV was used to test DNA samples from PBLs and milk samples of naturally infected cattle. Dr Christopher J. Finnegan from Veterinary Laboratories Agency (VLA) – Weybridge (Wielka Brytania) visited NVRI on June 20-26,.2009. The data showed that these primers succeed in spotting more positive samples than other others using DNA extracted from PBLs, however, DNA from milk cells was not suitable specimen for detection of DNA-BLV.
Genetic variability of env gene of BLV isolated from cattle in Ural region, Russia was the aim of bilateral collaborative study between reference laboratory at NVRI in Pulawy and Urals State Scientific Research Institute of Veterinary Medicine, in Ekaterinburg.
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
Szczotka M., Kuźmak J.: Identification of bovine dendritic cells phenotype and cytokine expression in cattle infected with BLV. 27th Meeting of ESVP/ECVP, 51, 09-12.09.2009, Kraków, Poland
Szczotka M., Kuźmak J.: Immunophenotype of bovine dendritic cells generated from monocytes of healthy and BLV infected animals. 3rd Annual Meeting EPIZONE “Crossing borders”, 12-15 May 2009, Antalya, Turkey,
¡ Other communications
A lecture “Infection with bovine leukemia virus: current diagnostic methods and molecular epidemiology” was presented on International Scientific-Practical Conference, Feodosia, Krym, Ukraina, 14-17 September, 2009.
Results of validation of a new format of ELISA test (SVANOVA) using panel of reference sera from cattle from Poland were collected and communicated to the supplier.
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?
N/A
ii) Did you submit to the OIE candidate kits for inscription on the OIE Register? If yes, for which kits?
N/A
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Annual reports of OIE Reference Laboratories and Collaborating Centres, 2009 3