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Home Biotechnologies Tests for Detection of Antibodies Against Diphtheria Toxin Subunits

Tests for Detection of Antibodies Against Diphtheria Toxin Subunits

Description

The diphtheria toxin contains two functional subunits: A (active) and B (binding). Each subunit carrying a distinct biological function implicated in the intoxication of the cell. It was shown in our studies that antibodies against A-subunit of diphtheria toxin (DT) prevalent over antibodies against Bsubunit of DT more often in blood serum of children with diphtheria as in blood serum of carriers or health vaccinated children. Therefore we propose to use immunologic test with recombinant A- and B-subunits as antigens for detection of antibodies against DT. Whole molecule of DT or toxoid (toxin after formaldehyde treatment) are used in others methods for this aim.

Variant 1. Immunoenzyme assay.
This assay allows determining quantity of antibodies against A- and B-subunits of DT in the generally accepted international units (IU). The level of antibodies against Bsubunit 0.1 IU or more is testifying to protectability against diphtheria. Therefore this assay may be used not only for diphtheria diagnosis, but for monitoring of protect antibodies’ level in population also.

Variant 2. Immunochromatography test.
Principal structure of immunochromatography test-system is shown on the picture. This test-system based on development of complexes antigen-antibodies by conjugate of protein A with colloidal gold. Results can be obtained for 10-15 minutes. Assay in variant 2 allow detecting a level of antitoxic antibodies only qualitatively. The intensity of band’s painting may be estimated quantitatively by densitometry.

Results of our laboratory investigations confirmed high specificity of developed test-systems and their correct work. We carried out investigation of antitoxic antibodies levels in blood serum of volunteers vaccinated against diphtheria. The obtained results verified possibility of clinical trials realization.

Innovative Aspect and Main Advantages

The main innovation of developed technology is separately using recombinant A- and B-subunits of diphtheria toxin as antigen for detection antibodies against diphtheria toxin. In others traditional technologies (RPGA or ELISA tests) whole molecule of diphtheria toxin or toxoid is usually used for this aim.

Key technical or competitive advantages by comparison with prevailing technologies:

  • Additional information about level of antibodies against each toxin subunit is important for diphtheria diagnosis;
  • Possibility to use test for broad screening investigations of immunity against diphtheria in human population;
  • Impossibility of influence therapeutic horse serum on the assay results;
  • High antigenic specificity of test due to more similarity of native diphtheria toxin to recombinant subunits than to anatoxin modified by formaldehyde;
  • Ease and rapidity of use;
  • Decreasing of assay value due to the fact that recombinant antigens are more chipper in comparison with native antigens.

Technical possibility to create others test-systems on the base developing approach.

Areas of Application

Immunodiagnosis of diphtheria, population investigations of immunity against diphtheria.


Contact Details

Dr. Kozulina Olena Petrivna
Palladin Institute of Biochemistry of NASU, Transfer Technology Office
Address: 9, Leontovicha str., 01601, Kyiv, Ukraine
(+38 044) 235 7157, Fax: (+38 044) 279 6365
This e-mail address is being protected from spambots. You need JavaScript enabled to view it

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