Main Article Content


K. Tursunov

National center of biotechnology, 13/5, Korgalzhyn rd., Astana, 010000, Kazakhstan

G. Raiymbek

National center of biotechnology, 13/5, Korgalzhyn rd., Astana, 010000, Kazakhstan

A.V. Shustov

National center of biotechnology, 13/5, Korgalzhyn rd., Astana, 010000, Kazakhstan

A. Begalieva

National center of biotechnology, 13/5, Korgalzhyn rd., Astana, 010000, Kazakhstan

B. Іnіrbay

National center of biotechnology, 13/5, Korgalzhyn rd., Astana, 010000, Kazakhstan

I. Sadiknabi

National center of biotechnology, 13/5, Korgalzhyn rd., Astana, 010000, Kazakhstan

K.K. Mukanov

National center of biotechnology, 13/5, Korgalzhyn rd., Astana, 010000, Kazakhstan

E.M. Ramanculov

National center of biotechnology, 13/5, Korgalzhyn rd., Astana, 010000, Kazakhstan

K.N. Mukantayev

National center of biotechnology, 13/5, Korgalzhyn rd., Astana, 010000, Kazakhstan


Bovine leukemia is an important infectious diseasethat affects farm animals. The viral etiology of the disease is well recognized. The disease causes significant economic damage to agricultural enterprises, including losses associated with the death and premature culling of high producing cows, reduced productivity, reduced milk quality, the cost of anti-leukemic measures. All these factors endanger the preservation of breeding herds, and threaten the management and breeding efforts to improve productive traits in dairy cattle.

In the present study, we determined that the optimal concentration of recombinant gp51 antigen to immobilize on a nitrocellulose membrane is 500 µg/ml. For the preparation of a colloidal gold conjugate, the optimal concentration of protein G is 8 µg/ml. The amount of colloidal gold conjugate for the membrane was 7 µl per 1 strip.

We found that the diagnostic characteristics of the immunochromatographic assay clearly differentiate the positive control sera of infected animals from sera of healthy animals, and also differentiates between serum samples with brucellosis and FMD. When determining the sensitivity of the test system, specific antibodies in the positive control sera were detected at a dilution of 1:1600.

Comparison of 145 positive serum samples in a lateral flow assay with immunoenzyme analysis and reaction immunodiffusion showed 97% agreement between the results.


virus, leukemia, gp51, recombinant antigen, immunochromatographic analysis, reaction diffusion precipitation

Article Details


Meas S., Ruas J., Farias N.A., Usui T. et al. Seroprevalence and molecular evidence for the presence of bovine immunodeficiency virus in Brazilian cattle. The Japanese journal of veterinary research, 2002, vol. 50, pp. 9-16. PMID: 12201018.

Kobayashi S., Tsutsui T., Yamamoto T., Hayama Y., Kameyama K., Konishi M., Murakami K.. Risk factors associated with within-herd transmission of bovine leukemia virus on dairy farms in Japan. Veterinary Research, 2010, vol. 6, pp. 1-9. PMID:20055982.

Coulston J., Naif H., Brandon R., Kumar S. et al. Molecular cloning and sequencing of an Australian isolate of proviral bovine leukaemia virus DNA: comparison with other isolates. Journal of General Virology, 1990, vol. 71, pp. 1737-1746. PMID: 2167927.

Burridge M.J., Puhr D.M., Hennemann J.M. Prevalence of bovine leukemia virus infection in Florida. Journal American Veterinary Medicine Associate, 1981, vol. 179, pp. 704-707. PMID:6281220.

Zhao X., Buehring G. Natural genetic variations in bovine leukemia virus envelope gene: possible effects of selection and escape. Virology, 2007, vol. 366, pp. 150-165. PMID:17498765.

Troiano L.D.C., Thomaz-Socco V., Agottani J.V.B., Brodzinski J. et al. Production, Characterization, and Use of Monoclonal Antibodies Against gp51 Protein to Diagnose Bovine Leukemia Virus Infection. BioResearch Open Access, 2013, vol. 2, no. 1, pp. 55- 60. doi: 10.1089/biores.2012.0295.

Ban J., Czene S., Altaner C., Callebaut I. et al. Mapping of sequential epitopes recognized by monoclonal antibodies on the bovine leukemia virus external glycoprotein expressed in Escherichia coli by means of antipeptide antibodies. Journal of General Virology, 1992, vol. 73, pp. 2457-2461. PMID: 1383413.

Siakkou H., Ulrich R., Uelze A., Mohring R., Rosenthal S. Immunological characterization of BLV proteins synthesized in Escherichia coli. Acta Virologica, 1990, vol. 34, pp. 256-262. PMID:1703391.

Ulrich R., Siakkou H., Platzer C., Bossmann H. et al. Synthesis of bovine leukemia virus antigen in Escherichia coli. Arch Exp Veterinarmed, 1990, vol. 44, pp. 909-916. PMID: 1966363.

Legrain M., Portetelle D., Dumont J., Burny A., Hilger F. Biochemical and immunological characterization of the bovine leukemia virus (BLV) envelope glycoprotein (gp51) produced in Saccharomyces cerevisiae. Gene, 1989, vol. 79, pp. 227-237. PMID: 2551774.

Kumar S., Andrew M.E., Boyle D.B., Brandon R.B. et al. Expression of bovine leukemia virus envelope gene by recombinant vaccinia virus. Virus Research, 1990, vol. 17, pp. 131-142.

Portetelle D., Limbach K., Burny A., Mammerickx M. et al. Recombinant vaccinia virus expression of the bovine leukemia virus envelope gene and protection of immunized sheep against infection. Vaccine, 1991, vol. 9, pp. 194-200. PMID: 1645899.

Kabeya H., Ohashi K., Ohishi K., Sugimoto C., Amanuma H., Onuma M. An effective peptide vaccine to eliminate bovine leukemia virus (BLV) infected cells in carrier sheep. Vaccine, 1996, vol. 14, pp. 1118-1122. PMID: 8911007.

Russo S., Montermini L., Berkovitz-Siman-Tov R., Ponti W., Poli G. Expression of bovine leukemia virus Env glycoprotein in insect cells by recombinant baculovirus. FEBS Lett, 1998, vol. 436, pp. 11-16. PMID: 9771885.

dos Santos E.M., Cardoso R., Filho L.R. G., Heinemann M.B. et al. Selection of ligand peptides with the ability to detect antibodies in enzootic bovine leucosis. African Journal of Biotechnology, 2012, vol. 11, pp. 7302-7312. doi: 10.5897/AJB11.348.

De Giuseppe A., Feliziani F., Rutili D., De Mia G.M. Expression of the bovine leukemia virus envelope glycoprotein (gp51) by recombinant baculovirus and its use in an enzyme linked immuno sorbent assay. Clinical and Diagnostic Laboratory Immunology, 2004, vol. 11, pp. 147-151. PMID:14715562.

Simard C., Richardson S., Dixon P., Belanger C., Maxwell P. Enzyme-linked immunosorbent assay for the diagnosis of bovine leukosis: comparison with the agar gel immunodiffusion test approved by the Canadian Food Inspection Agency. The Canadian Journal of Veterinary Research, 2000, vol. 64, pp. 101-106. PMID:10805248.

Peng D.P., Hu S.S., Hua Y., Xiao Y.C. et al. Comparison of a new gold-immunochromatographic assay for the detection of antibodies against avian influenza virus with hemagglutination inhibition and agar gel immunodiffusion assays. Veterinary Immunology and Immunopathology, 2007, vol. 117, pp. 17-25. doi:10.1016/j.vetimm.2007.01.022.

Byzova N.A., Zvereva E.A., Zherdev A.V., Eremin S.A., Dzantiev B.B. Rapid pretreatment free immunochromatographic assay of chloramphenicol in milk. Talanta, 2010, vol. 81, no. 3, pp. 843-848. doi: 10.1016/j.talanta.2010.01.025.

El-Eragi A.M., Salih M.H., Alawad M.F.E.M., Mohammed K.B. Evaluation of immunochromatographic assay for serodiagnosis of bovine brucellosis in Gezira State, Sudan. Veterinary World, 2014, vol. 7. Available at: doi: 10.14202/vetworld.2014.395-397.

Mi Young Park, Young Jin Kim, Sang Hyun Hwang, Hyoung Hoi Kim. et al. Chang. Evaluation of an immunochromatographic assay kit for rapid identification of Mycobacterium tuberculosis complex in clinical isolates. Journal of Clinical Microbiology, 2009, vol. 47, pp. 481-484. doi: 10.1128/JCM.01253-08.

Lau D.T., Ma H., Lemon S.M., Doo E. et al. A rapid immunochromatographic assay for hepatitis B virus screening. Journal of Viral Hepatitis, 2003, vol. 10, pp. 331-334. PMID: 12823602.