КОЛИЧЕСТВЕННАЯ ОЦЕНКА ПРОВИРУСНОЙ НАГРУЗКИ BLV У СИММЕНТАЛЬСКОГО КРС МЕТОДОМ ПЦР В РЕАЛЬНОМ ВРЕМЕНИ
DOI:
https://doi.org/10.52578/2305-9397-2025-2-2-98-110Ключевые слова:
лейкоз крупного рогатого скота, вирусная нагрузка, ПЦР (полимеразная цепная реакция) в реальном времени, гематологические ключи, BLV-инфекция (bovine leukosis virus)Аннотация
- Лейкоз крупного рогатого скота (ЛКРС), вызываемый вирусом лейкоза крупного рогатого скота (BLV), остаётся одной из ключевых проблем в молочном и мясном животноводстве. Особую эпидемиологическую опасность представляют субклинические формы инфекции, при которых животные длительно остаются вирусоносителями. В рамках настоящих исследований проведена количественная оценка провирусной нагрузки BLV методом полимеразной цепной реакции в режиме реального времени (ПЦР-РВ) у 46 голов симментальской породы. В качестве показателя использовалось отношение β-actin/env, отражающее уровень вирусной нагрузки. Результаты сопоставлены с гематологическими характеристиками животных, определёнными по ключам EC и JB. У особей с лимфоцитозом по JB Key (n = 8) медиана отношения β-actin/env составила 9,07, что было значительно ниже по сравнению с группой с нормальными показателями (медиана = 91; p = 0,0051), указывая на наличие высокой вирусной нагрузки. Кроме того, установлена статистически значимая обратная корреляция между результатами серологического теста (РИД) и отношением β-actin/env (ρ = –0,66; p < 0,001). Полученные данные подчёркивают диагностическую ценность комплексного подхода, включающего ПЦР-РВ, гематологический анализ и серологию, для эффективного выявления и контроля BLV-инфекции, включая бессимптомных носителей вируса
Библиографические ссылки
СПИСОК ЛИТЕРАТУРЫ
Heinecke, N., Tortora J., Martinez H.A., Gonzalez-Fernandez V.D. Detection and genotyping of
bovine leukemia virus in Mexican cattle // Archives of Virology. – 2017. – Vol. 162. –
P. 3191–3196.
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diagnosis of enzootic bovine leukosis // Zentralblatt für Veterinärmedizin Reihe B. – 1982. – Vol. 29. – P.
–680.
Martin, D. Comparative study of PCR as a direct assay and ELISA and AGID as indirect assays
for the detection of bovine leukaemia virus/ D. Martin [and etc.] // Journal of Veterinary Medicine Series
B. – 2001. – Vol. 48. – P. 97–106.
Buzala, E., Deren, W. Comparison of PLA with AGID and ELISA results in serology diagnosis
of bovine leukosis // Polish Journal of Veterinary Sciences. – 2003. – Vol. 6. – P. 9–11.
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leukemia cells forming colonies in the PHA-leukocyte feeder assay // Experimental Hematology. – 1981. – Vol. 9. – P. 588–594.
Andreolla, A.P., Erpen, L.M.S., Frandoloso, R. Development of an indirect ELISA based on
recombinant capsid protein to detect antibodies to bovine leukemia virus // Brazilian Journal of
Microbiology. – 2018. – Vol. 49, Suppl. 1. – P. 68–75. – DOI: 10.1016/j.bjm.2018.05.001.
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Vol. 71, № 9. – P. 2526–2534. – DOI: 10.3168/jds. S0022-0302(88)79841-0.
Nishiike, M., Haoka, M., Doi, T. Development of a preliminary diagnostic measure for bovine
leukosis in dairy cows using peripheral white blood cell and lymphocyte counts // Journal of Veterinary
Medical Science. – 2016. – Vol. 78, № 7. – P. 1145–1151. – DOI: 10.1292/jvms.16-0022.
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Mekata, H., Yamamoto, M., Kirino, Y. New hematological key for bovine leukemia virus
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P. 316–319. – DOI: 10.1292/jvms.17-0455.
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Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. – Available at:
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of bovine leukemia virus by loop-mediated isothermal amplification assay // Journal of Veterinary
Medical Science. – 2017. – Vol. 79, № 1. – P. 137–140. – DOI: 10.1292/jvms.16-0328.
Pereira, J.G. Diagnosis and phylogenetic analysis of bovine leukemia virus in dairy cattle in
northeastern Brazil // Frontiers in Veterinary Science. – 2022. – Vol. 9. – Article 1080994. – DOI:
3389/fvets.2022.1080994.
Kuckleburg, C.J., Chase, C.C., Nelson, E.A. Detection of bovine leukemia virus in blood and
milk by nested and real-time polymerase chain reactions // Journal of Veterinary Diagnostic Investigation. – 2003. – Vol. 15, № 1. – P. 72–76. – DOI: 10.1177/104063870301500117.
Kaur, R., Sodhi, M., Sharma, A. Selection of suitable reference genes for normalization of
quantitative RT-PCR (RT-qPCR) expression data across twelve tissues of riverine buffaloes (Bubalus
bubalis) // PLoS One. – 2018. – Vol. 13, № 3. – Article e0191558. – DOI: 10.1371/journal.pone.0191558.
Wang, G.H., Liang, C.C., Li, B.Z. Screening and validation of reference genes for qRT-PCR of
bovine skeletal muscle-derived satellite cells/ G.H Wang // Scientific Reports. – 2022. – Vol. 12, № 1. –
Article 5653. – DOI: 10.1038/s41598-022-09476-3.
Alvarez, I. Evaluation of total white blood cell count as a marker for proviral load of bovine
leukemia virus in dairy cattle from herds with a high seroprevalence of antibodies against bovine
leukemia virus // American Journal of Veterinary Research. – 2013. – Vol. 74, № 5. – P. 744–749. – DOI:
2460/ajvr.74.5.744.
Choi, K.Y., Monke, D., Stott, J.L. Absence of bovine leukosis virus in semen of seropositive
bulls // Journal of Veterinary Diagnostic Investigation. – 2002. – Vol. 14, № 5. – P. 403–406. – DOI:
1177/104063870201400507.
Cheng, T.Y., Zimmerman, J.J., Giménez-Lirola, L.G. Internal reference genes with the
potential for normalizing quantitative PCR results for oral fluid specimens // Animal Health Research
Reviews. – 2022. – Vol. 23, № 2. – P. 147–156.
Maksimova, V., Smith, S., Seth, J. HTLV-1 intragenic viral enhancer influences
immortalization phenotype in vitro, but is dispensable for persistence and disease development in animal
models // Frontiers in Immunology. – 2022. – Vol. 13. – Article 954077.
REFERENCES
Heinecke, N., Tortora J., Martinez H.A., Gonzalez-Fernandez V.D. Detection and genotyping of
bovine leukemia virus in Mexican cattle // Archives of Virology. – 2017. – Vol. 162. –
P. 3191–3196.
Bannenberg, T. Experiments using a modified disposition arrangement of the AGID test in the
diagnosis of enzootic bovine leukosis // Zentralblatt für Veterinärmedizin Reihe B. – 1982. – Vol. 29. – P.
–680.
Martin, D. Comparative study of PCR as a direct assay and ELISA and AGID as indirect assays
for the detection of bovine leukaemia virus/ D. Martin [and etc.] // Journal of Veterinary Medicine Series
B. – 2001. – Vol. 48. – P. 97–106.
Buzala, E., Deren, W. Comparison of PLA with AGID and ELISA results in serology diagnosis
of bovine leukosis // Polish Journal of Veterinary Sciences. – 2003. – Vol. 6. – P. 9–11.
Swart, K., Hagemeijer, A., Lowenberg, B. Density profiles and purification of chronic myeloid
leukemia cells forming colonies in the PHA-leukocyte feeder assay // Experimental Hematology. – 1981. – Vol. 9. – P. 588–594.
Andreolla, A.P., Erpen, L.M.S., Frandoloso, R. Development of an indirect ELISA based on
recombinant capsid protein to detect antibodies to bovine leukemia virus // Brazilian Journal of
Microbiology. – 2018. – Vol. 49, Suppl. 1. – P. 68–75. – DOI: 10.1016/j.bjm.2018.05.001.
Juliarena, M.A., Gutierrez, S.E., Ceriani, C. Determination of proviral load in bovine leukemia
virus-infected cattle with and without lymphocytosis // American Journal of Veterinary Research. – 2007. – Vol. 68, № 11. – P. 1220–1225. – DOI: 10.2460/ajvr.68.11.1220.
Ooshiro, M., Konnai, S. Horizontal transmission of bovine leukemia virus from lymphocytotic
cattle, and beneficial effects of insect vector control // The Veterinary Record. – 2013. – Vol. 173, № 21. – P. 527. – DOI: 10.1136/vr.101833.
Nikbakht Brujeni, G., Houshmand, P., Soufizadeh, P. Bovine leukemia virus: a perspective
insight into the infection and immunity // Iranian Journal of Veterinary Research. – 2023. – Vol. 24,
№ 4. – P. 290–300. – DOI: 10.22099/IJVR.2023.48236.7023.
Ruiz, V., Porta, N.G., Lomónaco, M. Bovine leukemia virus infection in neonatal calves. Risk
factors and control measures // Frontiers in Veterinary Science. – 2018. – Vol. 5. – Article 267. – DOI:
3389/fvets.2018.00267.
Lv, G., Wang, J., Lian, S. The global epidemiology of bovine leukemia virus: current trends and
future implications // Animals (Basel). – 2024. – Vol. 14, № 2. – Article 297. – DOI:
3390/ani14020297.
Lewin, H.A., Wu, M.C., Nolan, T.J. Peripheral B lymphocyte percentage as an indicator of
subclinical progression of bovine leukemia virus infection // Journal of Dairy Science. – 1988. – Vol. 71,
№ 9. – P. 2526–2534. – DOI: 10.3168/jds. S0022-0302(88)79841-0.
Nishiike, M., Haoka, M., Doi, T. Development of a preliminary diagnostic measure for bovine
leukosis in dairy cows using peripheral white blood cell and lymphocyte counts // Journal of Veterinary
Medical Science. – 2016. – Vol. 78, № 7. – P. 1145–1151. – DOI: 10.1292/jvms.16-0022.
Bendixen, H.J. Preventive measures in cattle leukemia: leukosis enzootica bovis // Annals of the
New York Academy of Sciences. – 1963. – Vol. 108. – P. 1241–1267. – DOI: 10.1111/j.1749-6632.
tb13448. x.
Da, Y., Shanks, R.D., Stewart, J.A. Milk and fat yields decline in bovine leukemia virus
infected Holstein cattle with persistent lymphocytosis // Proceedings of the National Academy of
Sciences of the United States of America. – 1993. – Vol. 90, № 14. – P. 6538–6541. – DOI:
1073/pnas.90.14.6538.
Mekata, H., Yamamoto, M., Kirino, Y. New hematological key for bovine leukemia virus
infected Japanese Black cattle // Journal of Veterinary Medical Science. – 2018. – Vol. 80, № 2. –
P. 316–319. – DOI: 10.1292/jvms.17-0455.
OIE (World Organization for Animal Health). Chapter 3.4.9. Enzootic Bovine Leucosis //
Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. – Available at:
www.woah.org/fileadmin/Home/fr/Health_standards/tahm/3.04.09_EBL.pdf
Okuwa, Y., Miyamato-Hayashi, M., Tanaka, T. Simple and rapid method for routine screening
of bovine leukemia virus by loop-mediated isothermal amplification assay // Journal of Veterinary
Medical Science. – 2017. – Vol. 79, № 1. – P. 137–140. – DOI: 10.1292/jvms.16-0328.
Pereira, J.G. Diagnosis and phylogenetic analysis of bovine leukemia virus in dairy cattle in
northeastern Brazil // Frontiers in Veterinary Science. – 2022. – Vol. 9. – Article 1080994. – DOI:
3389/fvets.2022.1080994.
Kuckleburg, C.J., Chase, C.C., Nelson, E.A. Detection of bovine leukemia virus in blood and
milk by nested and real-time polymerase chain reactions // Journal of Veterinary Diagnostic Investigation. – 2003. – Vol. 15, № 1. – P. 72–76. – DOI: 10.1177/104063870301500117.
Kaur, R., Sodhi, M., Sharma, A. Selection of suitable reference genes for normalization of
quantitative RT-PCR (RT-qPCR) expression data across twelve tissues of riverine buffaloes (Bubalus
bubalis) // PLoS One. – 2018. – Vol. 13, № 3. – Article e0191558. – DOI: 10.1371/journal.pone.0191558.
Wang, G.H., Liang, C.C., Li, B.Z. Screening and validation of reference genes for qRT-PCR of
bovine skeletal muscle-derived satellite cells/ G.H Wang // Scientific Reports. – 2022. – Vol. 12,
№ 1. – Article 5653. – DOI: 10.1038/s41598-022-09476-3.
Alvarez, I. Evaluation of total white blood cell count as a marker for proviral load of bovine
leukemia virus in dairy cattle from herds with a high seroprevalence of antibodies against bovine
leukemia virus // American Journal of Veterinary Research. – 2013. – Vol. 74, № 5. – P. 744–749. – DOI:
2460/ajvr.74.5.744.
Choi, K.Y., Monke, D., Stott, J.L. Absence of bovine leukosis virus in semen of seropositive
bulls // Journal of Veterinary Diagnostic Investigation. – 2002. – Vol. 14, № 5. – P. 403–406. – DOI:
1177/104063870201400507.
Cheng, T.Y., Zimmerman, J.J., Giménez-Lirola, L.G. Internal reference genes with the potential
for normalizing quantitative PCR results for oral fluid specimens // Animal Health Research Reviews. –
– Vol. 23, № 2. – P. 147–156.
Maksimova, V., Smith, S., Seth, J. HTLV-1 intragenic viral enhancer influences
immortalization phenotype in vitro, but is dispensable for persistence and disease development in animal
models // Frontiers in Immunology. – 2022. – Vol. 13. – Article 954077.
