Genotyping of BLV isolates spread at the territory of the Republ

Gulyukin M.I., Kozyreva N.G., Gendzhieva O.B., Ivanova L.A.

Summary

Amplification of the desired fragment of the pol gene was carried out with the use of developed primers PF2-PR2 complementary to a section of the BLV pol gene. The 438 bp fragments of the genome of BLV provirus isolates from mature seropositive cows were obtained by the PCR method. The nucleotide sequence of a section of the provirus pol gene of bovine leukemia virus (BLV) isolates from animals from some farms in Gorodovikovsky region of the Republic of Kalmykiya is determined whereby permit concluding homology of circulating viruses and belong to the same group as BLV isolates from Argentina, Australia, Brazil, and the United States. On the basis of the revealed homology of the section of the pol gene of BLV isolates, primers PF2-PR2 can be used also for detecting BLV proviral DNA when conducting routine PCR diagnosis of bovine leucosis.

Key words: bovine leucosis virus, PCR, sequencing, phylogenetic analysis.

References

1. Kozyreva N.G., Gulyukin M.I., Ivanova L.A., Malogolovkin A.S., Klimenko A.I., Razumovskaya V.V. Filogenetichesky analiz uchastka gena pol izolyatov provirusa leykoza KRS, obnaruzhennykh u zhivotnykh iz razlichnykh hozyaystv regionov Rossiyskoy Federatsii [Phylogenetic analysis of pol gene site of leucosis provirus isolates found in animals from different farms of regions of Russia]. – 2011. – pp. 48-50.
2. Limansky A.P., Limanskaya O.Yu. Molekulyarno-geneticheskie metody – osnova programm po iskoreneniyu leykoza krupnogo rogatogo skota [Molecular genetic methods – basis of eradication programs of bovine leucosis]. – Biotehnologiya, 2001 (#3):40-50.
3. Ponomareva I.S., Sycheva M.V. Dinamika infitsirovannosti, biologichesky b immunologichesky testy pri leykoze korov na Yuzhnom Urale [Dynamics of infection rates, biochemical and immunological tests for leucosis of cows in the southern Urals]. – Veterinarnaya patologiya, 2010, #2. – pp. 69-71.
4. Porodny sostav krupnogo rogatogo skota v hozyaystvakh Kalmytskoy ASSR [Species composition of cattle on farms of the Kalmyk ASSR]. – 1982. – 59 p.
5. Smirnov P.N., Nezavitin A.G., Smirnova V.V., Khramtsov V.V. Problemy leykoza zhivotnykh [Problems of animal leucosis]. – Novosibirsk, 1992. – 468 p.
6. Prokhvatilova L.B., Lomakin A.I., Kolosov S.N., Drygin V.V., Rybakov S.S., Gusev A.A. Diagnostika leykoza KRS metodom polimeraznoy tsepnoy reaktsii [Diagnostics of bovine leucosis by polymerase chain reaction]. – 1998 (#5). – pp. 65-68.
7. Asfaw Y. Distribution and superinfection of bovine leukemia virus genotypes in Japan/ Tsuduku S., Konishi M. et al. // Arch. Virol. – 2005. – N150. – Р. 493-505.
8. Dube S., Abbott L., Dube D.K., Dolcini G., Gutierrez S., Ceriani C., Juliarena M., Ferrer J., Perzova R., Poiesz B. J. The complete genomic sequence of an in vivo low replicating BLV strain // J. Virol. – 2009. – N6:120. (URL: http://www.virologyj.com/content/6/1/120).
9. Fechner H., Blankenstein P., Looman A.C. Provirus Variants of the Bovine Leukemia Virus and Their Relation to the Serological Status of Naturally Infected Cattle // J.Virol. – 1997. – N237. – P.261-269.
10. Felmer R. G. Molecular analysis of a 444 bp fragment of the BLV gp51 env gene reveals a high frequency of non-silent point mutations and suggest the presence of two subgroups of BLV in Chile // J. Vet. Microb. – 2005. – N108. – P. 39-47.
11. Gillet N., Florins A., Boxus M. C. Burteau, Nigro A., Vandermeers F., Balon H., Bouzar A.-B., Defoiche J., Burny A., Reichert M., Kettmann R., Willems L. Mechanisms of leukemogenesis induced by bovine leukemia virus: prospects for novel anti-retroviral therapies in human// J. Retrovirol. – 2007. – N4. – P.18-50.
12. Licursi M., Inoshima Y., Wu D., Yokoyama T., González E.T., Sentsui H. Genetic heterogeneity among bovine leukemia virus genotypes and its relation to humoral responses in hosts // Virus Res. – 2002. – N86. – P.101- 110.
13. Rodriguez S.M., Golemba M.D., Campos R.H., Trono K., Jones L.R. Bovine leukemia virus can be classified into seven genotypes: evidence for the existence of two novel clades // J. Gen.Virol. – 2009. – N90.– P.2788–2797.

Author affiliation

Gulyukin Mikhail I., D.Sc. in Veterinary Medicine, professor, academician of the Russian Academy of Agricultural Sciences, director of the Kovalenko All-Russian Institute of Experimental Veterinary Medicine; 24/1, Ryazansky ave., Moscow, 109428; phone: 8(495)9700368; e-mail: viev@mail.ru.

Ivanova Lyudmila A., Ph.D. in Biology, leading scientific researcher of the laboratory of leucosis of the Kovalenko All-Russian Institute of Experimental Veterinary Medicine; 24/1, Ryazansky ave., Moscow, 109428; phone: 8(495)9700366; e-mail: viev@mail.ru.

Gendzhieva Olga B., Ph.D. in Veterinary Medicine, senior lecturer of the department of zootechny of the Kalmyk state university; phone: 8(847)2239038; e-mail: gend_olga@mail.ru.

Responsible for correspondence with the editorial board:

Kozyreva Nataliya G., junior scientific researcher of the laboratory of leucosis of the Kovalenko All-Russian Institute of Experimental Veterinary Medicine; 24/1, Ryazansky ave., Moscow, 109428; phone: 8(495)9700366, 8-915-113-75-00; e-mail: nk07-73@rambler.ru.