Preview

Problems of Particularly Dangerous Infections

Advanced search

Studies of the Composition of the Cell Antigen Preparations Obtained from Pathogenic Burkholderia, Using Immunoblotting

https://doi.org/10.21055/0370-1069-2015-2-46-49

Abstract

Carried out has been comparative analysis of the cell antigen immune-electrophoregrammes of B. pseudomallei C-141, B. mallei 10230, B. thailandensis 264, and B. cepacia 25416 after immunoblotting with immune rabbit sera to living cells of avirulent B. pseudomallei 107, celllular and extracellular antigens of B. pseudomallei 107, B. pseudomallei C-141, B. mallei 10230, B. thailandensis 264, and B. cepacia 25416. Visual investigation has revealed the presence of fractions with molar mass of 18.4 and 35 kDa in B. malei 10230 cell antigens’ electrophoregramme obtained by means of immunoblotting with serum to B. malei 10230 cell antigens, as distinct from B. pseudomallei C-141 electrophoregramme with the same serum. This makes it possible to distinguish the pathogenic Burkholderia. At the same time utilization of heterologous immune sera enhances discriminating capacity of the comparative assay. The level of reproducibility of immune-electrophoregrammes allows for the deployment of the similarity coefficient for computer-based comparative analysis.

About the Authors

A. A. Budchenko
Volgograd Research Anti-Plague Institute
Russian Federation


I. Yu. Mazurova
Volgograd Research Anti-Plague Institute
Russian Federation


V. I. Ilyukhin
Volgograd Research Anti-Plague Institute
Russian Federation


References

1. Beily N. [Mathematics in Biology and Medicine]. M.: Mir; 1970.

2. Budchenko A.A., Ilyukhin V.I., Viktorov D.V. [Comparative analysis of electrophoregrammes of bulk cell proteins in pathogenic Burkholderia]. Mol. Genet. Mikrobiol. Virusol. 2005; 2:24–8.

3. Domaradsky I.V. [Problems of Cross-Protective Immunity]. M.: Meditsina; 1973.

4. Ilyukhin V.I., Senina T.V., Plekhanova N.G., Antonov V.A., Merinova L.K., Seimova I.K. [Burkholderia thailandensis: biological properties, identification and taxonomic position]. Mol. Genet. Mikrobiol. Virusol. 2002; 1:7–11.

5. Kulakov Yu.K., Zheludkov M.M., Tolvacheva T.A. [Application of molecular-genetic methods for Brucella identification in comparative assay of the strains isolated from infected dogs]. Mol. Genet. Mikrobiol. Virusol. 2003; 1:6–14.

6. Anuntagool N., Rugdech P., Sirisinha S. Identification of specific antigens of Pseudomonas pseudomallei and evaluation of their efficacies for diagnosis of melioidosis. J. Clin. Microbiol. 1993; 31(1):1232–6.

7. Bradford M.M. A rapid and sensitive method for the guantitation of microgram guantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 1976; 72:248–54.

8. Katz J., Devvald R., Nicholson J. Procedurally similar competitive immunoassay systems for the serodiagnosis of Babesia eaui, Babesia eaballi, Trypanosoma euuiperdum and Burkhohleria mallei infection in horses. J. Vet. Diagn. Invest. 2000; 12:46–50.

9. Laemmli U.K. Clevage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227: 680–5.

10. Pal V., Kumar S., Malik P., Rai G.P. Evaluation of Recombinant Proteins of Burkholderia mallei for Serodiagnosis of Glanders Clin. Vaccine Immunol. 2012; 19:1193–8.

11. Sermswan R.W., Wongratanacheewin S., Anuntagool N. Comparison of the polymerase chain reaction and serologic tests for diagnosis of septicemic melioidosis. J. Trop. Med. Hyg. 2000; 63:146–9.

12. Tiyawisutsri R., Peacock S.J., Langa S., Limmathurotsakul D., Cheng A.C., Chierakul W., Chaowagul W., Day N.P., Wuthiekanun V. Antibodies from patients with Melioidosis recognize Burkholderia mallei but not Burkholderia thailandensis antigens in the indirect hemagglutination assay. J. Clin. Microbiol. 2005; 43:4872–4.

13. 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; 76:4350–4.


Review

For citations:


Budchenko A.A., Mazurova I.Yu., Ilyukhin V.I. Studies of the Composition of the Cell Antigen Preparations Obtained from Pathogenic Burkholderia, Using Immunoblotting. Problems of Particularly Dangerous Infections. 2015;(2):46-49. (In Russ.) https://doi.org/10.21055/0370-1069-2015-2-46-49

Views: 527


Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.


ISSN 0370-1069 (Print)
ISSN 2658-719X (Online)