Main Article Content


A.K. Bulashev

S. Seifullin Kazakh Agro-Technical University, 2 Altynsarin Str., Astana, 010011, Kazakhstan

Zh.A. Suranshiev

S. Seifullin Kazakh Agro-Technical University, 2 Altynsarin Str., Astana, 010011, Kazakhstan

A.Kh. Zhumalin

S. Seifullin Kazakh Agro-Technical University, 2 Altynsarin Str., Astana, 010011, Kazakhstan

K.А. Tursunov

S. Seifullin Kazakh Agro-Technical University, 2 Altynsarin Str., Astana, 010011, Kazakhstan


The effectiveness of brucellosis eradication measures depends on the timely diagnosis and isolation of infected animals. Commercial ELISA-kits for diagnosis of brucellosis use lipopolysaccharides (LPS) as target antigens. It is well known that use of these cell wall components can result in false-positive results due to cross-reaction of antibodies against Brucella spp. with other gram-negative bacteria. Outer membrane proteins (OMP) are of great interest, as they are specific, not only to the Brucella genus, but also for particular species. OMP extracted from Brucella abortus 19 and Brucella melitensis Rev-1 were more antigenic than LPS by agglutination and complement fixation tests using serum samples from cows positive for brucellosis. B. abortus OMP also had significantly greater diagnostic value than an antigenic recombinant protein with a molecular weight of 26 kDa (rBP26). A noticeable advantage of OMP over rBP26 was also established by serological investigation of cows vaccinated with the rough strain, B. abortus RB-51. The results suggest that in the development of ELISA-kits for diagnosis of bovine brucellosis not a single protein, but several recombinant proteins, with distinct antigenic properties should be used. Among OMP protein fractions from both Brucella species, those with the greatest potential as diagnostic antigens had molecular weights of 19 and 15 kDa, in addition to a B. melitensis protein, with a molecular weight of 12 kDa. These proteins demonstrated antigenicity in western blot analysis using serum antibodies from cows positive for brucellosis by serological tests and PCR.


brucellosis, Brucella abortus, Brucella melitensis, outer membrane proteins, antigenicity, ELISA

Article Details


Kisykov T. Epidemiological situation in Kazakhstan brucellosis cattle. Veterinary (Kaz.),2009, vol. 3(7), pp. 44-46.

Ivanov N.I. Brucellosis. J.AgroӘlem,2010, vol.2(7), pp.40-47.

Plotnikova A.M., Salmakov K.M., Ivanov A.V. Immune monitoring of animal brucellosis. J. Veterinary (RF),2010, vol.5, pp. 26-30.

Bulashev A.K., Suranshiev Z. Using Brucella abortus outer membrane proteins in serodiagnosis of Brucellosis. Abstracts of Eight Annual International Symposium on Agriculture, Third Annual International Conference on Ecology, Ecosystems and Climate Change & Third Annual International Forum on Water, Athens, Greece, July 2015, pp.43-44.

Bundle D.R., Gidney M.A., Perry M.B., Duncan J.R., Cherwonogrodzky J.W.Serological confirmation of Brucella abortus and Yersinia enterocolitica O:9 O-antigens by monoclonal antibodies. Infection and immunity,1984, vol.46 (2), pp. 89-93.PMID: 6437982.

Corbel M.J., Stuart F.A., Brewer R.A. Observations on serological cross – reactions between smooth Brucellaspecies and organisms of other genera. Dev.Biol.Stand., 1984, vol. 56, pp. 341-405.PMID: 6489620.

Corbel M.J. Recent Advances in the study of Brucella antigens and their serological cross reactions. Vet. Bull., 1985, vol.55, no. 12, pp. 927-942.

Gupta V.K., Verma D.K., Singh S.V., VihanV.S. Serological diagnostic potential of recombinant outer membrane protein (Omp31) from Brucella melitensisin goat and sheep brucellosis.Small Ruminant Research,2007, vol. 70, no. 2-3, pp. 260-266.

Lim J.J.,Kim D.H.,Lee J.J.,Kim D.G.,Min al. Evaluation of recombinant 28 kDa outer membrane protein of Brucella abortus for the clinical diagnosis of bovine brucellosis in Korea.J.Vet. Med. Sci., 2012, vol. 74, no. 6, pp. 687-691. PMID: 22214857.

Ko K.Y., Kim J., Her M. et al. Immunogenic proteins of Brucella abortus to minimize cross reactions in brucellosis diagnosis.Vet. Microbiology, 2012, vol.156, no. 3-4, pp.374-380. doi: 10.1016/j.vetmic.2011.11.011.

Patent №14230 Republic of Kazakhstan, G01N 33/535.A method of detecting antibodies against the causative agent of brucellosis.Shenzhanov K.T., SuranshievZh.A., Bulashev A.K., Ospanova S.G.; statement22.07.2002, published 15.04.2008, bulletin 4, p.4.

Bradford M. A rapid and sensitive methodfer the qvantitation of microgram qvantitaties of protein utilizing the principle of protein – due binding.Anal.Biochem., 1976, vol. 72, no. 4, pp.248-254.

Laemmli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature,1970, vol.227, pp.680-685.PMID: 5432063.

Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci., USA,1979, vol.76, no. 9, pp.4350-4354.PMID: 1422008.

Sayduldin T.C. Statistical analysis of the results of serological tests.J. Veterinary, 1981, no. 7, pp.62-66.

Cassataro J.,Pasquevich K.,Bruno L.,Wallach J.C.,Fossati C.A.,Baldi P.C. Antibody reactivity to Omp31 from Brucella melitensis in human and animal infections by smooth and rough Brucellae.Clin.Diagn. Lab. Immunol.,2004, vol. 11, no. 1, pp.111-114.PMID: 14715555.

Simborio H.L. ,Lee J.J.,Bernardo Reyes A.W.,Hop H.T.,Arayan L.T., Min W. et al. Evaluation of the combined use of the recombinant Brucella abortus Omp10, Omp19 and Omp28 proteins for the clinical diagnosis of bovine brucellosis. Microb.Pathog.,2015, vol. 83/84, pp. 41-46. doi: 10.1016/j.micpath.2015.05.004.

Tibor A., Saman E.,Wergifosse P. de, Cloeckaert A.,Limet J.N., Letesson J.J. Molecular characterization, occurrence, and immunogenicity in infected sheep and cattle of two minor outer membrane proteins of Brucella abortus.Infect Immun., 1996,vol. 64(1), pp.100-107. PMID: 8557326.