New recombinant T. pallidum antigens Tp0453 and Tp0319 in the diagnostics of syphilis

Cover Page

Abstract


A new technology for obtaining a recombinant version of a new dedicated protein T. pallidum Tp0319 for the serological diagnostics of syphilis was gene engineered. As a result, a homogeneous (according to denaturating polyacrylamide gel electrophoresis) recombinant version of T. pallidum Tp0319 protein was obtained. The molecular mass of recombinant T. pallidum Tp0319 protein according to electrophoretic mobility is about 37 kDa, which corresponds to the molecular mass of a fragment of Tp0319 protein without a signal peptide calculated based on the amino-acid sequence (37.3 kDa). According to the developed technology, purified recombinant T. pallidum Tp0319 and Tp0453 proteins were obtained. The proteins were used as antigens as a part of a solid-phase immunoadsorbent for detecting specific IgG to the study protein in the serum of patients suffering from different types of acquired syphilis. The use of the resulting recombinant protein enabled the authors to reveal T. pallidum antibodies in the blood serum in patients suffering from syphilis (primary, secondary, latent early and late stage syphilis). The examination of blood serum samples taken from healthy donors revealed no antibodies. Based on the study results, it is possible to make a conclusion about the possibility to use the resulting recombinant protein (Тр0319) as an extra antigen for diagnostics of syphilis. The introduction of the new antigen in the immunoadsorbent for test systems used for diagnosing syphilis (in the form of immune-enzyme assay, immunoblotting or immune chips) expands the potential of serological diagnostics of this disease due to the expansion of the range of T. pallidum antibodies to be revealed.

A. V. Runina

State Research Center of Dermatovenereology and Cosmetology, Ministry of healthcare of the Russian Federation

Author for correspondence.
Email: runina@cnikvi.ru

Russian Federation

R. F. Khairullin

State Research Center of Dermatovenereology and Cosmetology, Ministry of healthcare of the Russian Federation

Email: noemail@neicon.ru

Russian Federation

K. V. Rog

State Research Center of Dermatovenereology and Cosmetology, Ministry of healthcare of the Russian Federation

Email: noemail@neicon.ru

Russian Federation

V. I. Semina

State Research Center of Dermatovenereology and Cosmetology, Ministry of healthcare of the Russian Federation

Email: noemail@neicon.ru

Russian Federation

S. V. Rotanov

State Research Center of Dermatovenereology and Cosmetology, Ministry of healthcare of the Russian Federation

Email: noemail@neicon.ru

Russian Federation

  1. Chi B., Chauhan S., Kuramitsu H.; Development of a System for Expressing Heterologous Genes in the Oral Spirochete Treponema denticola and Its Use in Expression of the Treponema pallidum flaA Gene. Inf immun 1999; 67 (7): 3653-3656.
  2. Cameron C.E., Kuroiwa J.M., Yamada M., Francescutti T., Chi B., Kuramitsu H.K. Heterologous expression of the Treponema pallidum lamininbinding adhesin Tp0751 in the culturable spirochete Treponema phagedenis. J bacteriol 2008; 190 (7): 2565-2571. Литература
  3. Khairullin R.F., Rotanov S.V., Frigo N.V., Belousova A.V. Bioinformatic analysis of T. pallidum specific antigens Vestn Dermatol Venerol 2012; 5: 56-64.
  4. Cox D.L., Luthra A., Dunham-Ems S., Desrosiers D.C., Salazar J.C., Caimano M.J., Radolf J.D. Surface immunolabeling and consensus computational framework to identify candidate rare outer membrane proteins of Treponema pallidum. Inf immun 2010; 78 (12): 5178-5194.
  5. Laemmli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. nature 1970; 227 (5259): 680-685.
  6. Bradford M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical biochemistry 1976; 72 (1): 248-254.
  7. Khairullin R.F., Runina A.V., Rog K.V., Frigo N.V., Rotanov S.V. Production and evaluation of the clinical efficacy of the recombinant T. pallidum antigen Tp0453 for syphilis diagnosis. Vestn Dermatol Venerol 2013; 6: 73-79.

Views

Abstract - 92

PDF (Russian) - 32

Refbacks

  • There are currently no refbacks.

Copyright (c)



This website uses cookies

You consent to our cookies if you continue to use our website.

About Cookies