ANTIGENICITY OF BRUCELLA PROTEINS BY THE ELISA TEST

Lifetime diagnostics of animal brucellosis is mainly based on serological reactions as SAT, RBPT and CFT. The tests determine antibodies by means of antigen produced from Brucella S-cells that mainly contain lipopolysaccharides (LPS). The LPS may cause cross-reactions with other clinically significant gram-negative bacteria; this leads to false-positive results. Due to this fact, the researchers involved in improving. The paper highlights the research results on antigenicity of 5 recombinant Brucella proteins (rOMP19, rOMP25, rOMP31, rBP26 and rSOD) and soluble protein preparations (CSP) of B. abortus and/or B. melitensis by indirect ELISA using cattle and sheep serum samples positive for brucellosis by classical serological tests. CSP appeared to be the most antigenic among the protein specimens; it determined antibodies in 94.8% of the cattle and 69% of sheep. Antibodies which were specific to rOMP19, rOMP25 and rOMP31 were detected in 39%; 50.6 and 76.6% of antibody-positive cows. Periplasmic proteins (rBP26 and rSOD) were observed as less antigenic than outer membrane proteins and revealed anti-Brucella antibodies in 29.9 and 14.3% of the cattle. Recombinant proteins were not detected by antibodies of sheep positive for brucellosis. Antibodies to recombinant proteins by i-ELISA were detected in the small number of the cattle kept at brucellosis free farm (from 2.1 to 12.5%). The results obtained outline the necessity to carry out experimental infection of animals in order to assess properly the capacities of recombinant proteins when diagnosing brucellosis.

1. Pappas G., HCDCP E-bulletin, available at: http://www2.keelpno.gr/blog/?p=2033

2. Arakelyan P.K., Raznitsina G.V., Yanchenko T.A., Bondarev Ye.G., Dimova A.S., Dimov S.K., Chekishev V.M., Vorob’yev V.I. Veterinariya, 2016, No. 7, pp.25–30. (In Russ.)

3. Baldi P.C., Giambartolomei G.H., Goldbaum F.A., Abdón L.F., Velikovsky C.A., Kittelberger R., Fossati C.A. Clinical and Diagnostic Laboratory Immunology, 1996. Vol. 3, pp.472–476.

4. Eshmuhametov A.E., Bejsembaev K.K., Asauova Zh.S., Sultanova A.O. Vestnik Kostanajskogo gosudarstvennogo universiteta im. A. Bajtursunova, 2016, No. 2, pp.36–41. (In Russ.)

5. Ko K.Y., Kim J.W., Her M., Kang S.I., Jung S.C., Cho D.H., Kim J.Y., Veterinary Microbiology, 2012, Vol. 156, pp.374–380.

6. Kaur G., Verna R., Kumar B.V.S., Deka D., Arawall R.K. Proceedings of the National Academy of Sciences, 2015. Vol.85, pp.853–858.

7. Mohammadi E., Golchin M. Adv Clin Exp Med., 2018, Vol. 27, pp.643–648.

8. Cloeckaert A., Verger J.M., Grayon M., Zygmunt M.S., Grépinet O. Infect. Immun, 1996, Vol.64. pp. 2047–2055.

9. Navarro-Soto M.C., Gomez-Flores R., Morales- LoredoA., Ramírez-Pfeiffer C., Tamez-Guerra P., ÁlvarezOjeda G., Biotechnology Summit, Proceedings of the Conference, 8–10 Oct., 2014, Mexico, pp.139–143.

10. Manat Y., Shustov A.V., Evtehova E., Eskendirova S.Z. Open Veterinary Journal, 2016, Vol. 6, pp. 71–77.

11. PrattA.J., DiDonato M., Shin D.S., Cabelli D.E., Bruns C.K., Belzer C.A., GorringeA.R., Langford P.R., Tabatabai L.B., Kroll J.S., Tainer J.A., Getzoff E.D. Bacteriol, 2015, Vol. 197, pp.3834–3847.

12. Tabatabai L.B., Deyoe D.L. Developments in biological standardization, Vol. 56, pp.199–211.

13. Bulashev A.K., Tursunov K.T., Kairova Zh.K. Vestnik KazATU im. S. Seifullina, 2018, No. 3, pp. 117– 127. (In Russ.)

14. Eskendirova S.Z., Shustov A.V., Evtyhova E.B., Manat E., Patent RK № 30873., 2014, byul. No. 12, 5 p. (In Russ.)

15. Ahmed I.M., Khairani-Bejo S., Hassan L., Bahaman A.R., Omar A.R., BMC Vet. Res., 2015, Vol. 11: 275. DOI 10.1186/s12917–015–0587–2.