[IDENTIFICATION OF CAUSATIVE AGENTS OF GLANDERS AND MELIOIDOSIS BASED ON PRINCIPLES OF POLYPHASE TAXONOMIC APPROACH].

AIM: Determine an optimal set of the most effective methods of identification and intraspecies typing ofcausative agents ofglanders and melioidosis. Materials andmethods. Bacteriologic, immunochemical, molecular-genetic methods were used. RESULTS: A possibility to identify collection strains of pathogenic and closely related Burkholderia in semiautomatic systems is studied. Means of detection of informative variable genome segments ofthe specified microorganisms were developed, methods of their genetic typing were selected. Effectiveness of application of precipitating mAbs for differentiation of Burkholderia was established. Data on diagnostic possibilities of immunoglobulins fluorescing based on monoclonal antibodies of various etiotropic directionality for detection and identification of B. mallei and B. pseudomallei are generalized. Experimental series of amplification test-systems for identification of glanders and melioidosis causative agents in real-time PCR format are created. CONCLUSION: A number of methods for identification and typing of glanders and melioidosis causative agents is proposed.

[The comparative evalution of informativeness of immunologic and molecular genetic methods and means during stages of specific indication of melioidosis agent].

The reference-center of monitoring of agents of glanders and melioidosis carried out testing of reagents kits for diagnostic of agent of melioidosis and other close-related species of Burkholderiae in vitro. At the stage of specific identification of pathogenic Burkholderiae the diagnostic possibilities of commercial and experimental kits of reagents for express- and rapid analysis were evaluated. The criteria of evaluation of diagnostic value of kits of reagents were sensitivity, specificity and time of implementation of studies. The analysis with application of mono- and multi-locus amplification systems, including real-time polymerase chain reaction permitted during 5-6 hours to implement identification and differentiation of Burkholderia pseufomallei, B. thailandensis and B. cepacia.

[Immunogenic and protective characteristics of surface antigens 6 and d of Burkholderia pseudomallei].

AIM: Extraction of complex of Burkholderia pseudomallei antigens 6+d (Ag6+d) and study of its immunogenic and protective characteristics. MATERIALS AND METHODS: Studied antigens were obtained from acrtone-dried cells of B. pseudomallei 57576. Experiments were performed on white mouse model. Microbiological, immunochemical as well as immunological methods were used in the study. RESULTS: It was shown that antigenic complex 6+d has a glycoprotein nature and corresponds to high-polymeric catode-moving antigen 6 with molecular mass 500 kDa and anode-moving antigen d with molecular mass 40 - 55 kDa. Immunogenic and protective characteristics of surface antigenic complex 6+d was studied. It was noted that Ag6+d caused reliable stimulating effect on cellular arm of immune system of white mice appeared in activation of delayed-type hypersensitivity reactions, enhanced phagocytic activity of polymorphonuclear macrophages as well as increased expression of Fc-receptors on macrophages and neutrophils. CONCLUSION: Observed stimulation of cellular immunity by surface antigens was confirmed during study of their protective characteristics on the model of experimental meloidosis in white mice.

[Immunogenicity of surface and capsular antigens of Burkholderia].

Study showed that five (C3, C6, C9, C10, C11) out of ten chromatographic fractions of surface and capsular antigens of B. mallei significantly stimulated cell-mediated immunity that manifested in activation of delayed hypersensivity reactions (DHS) and phagocyteability of noncapsulated avirulent strain of B. mallei with added surface and capsular antigenic complexes. Other fractions did not stimulate cell-mediated immunity, furthermore, fraction C8, which contained capsular biopolymer with mass of 200 kD (Ar8), was characterized by immunosuppressive effect on DHS and phagocytosis. Observed stimulation of cell-mediated immunity by fractions referred above has been confirmed by assessment of their protective effects on the model of experimental melioidosis in white rats. Relationship between markers of humoral and cell-mediated immunity, including markers of specific response, was not observed.

[Role of toxoplasmas in pathology of the vision organ].

The present paper presents data on the major clinical symptoms ofToxoplasma oculopathy in different forms of Toxoplasma infection. It also gives the results of the authors' own laboratory studies of specimens taken from individuals with pathology of the vision organ. They confirm the data available in the literature on that toxoplasmosis frequently occurs in combination with other infections, particularly with Herpes and Cytomegalovirus, Chlamydia, Ureaplasma, and Mycoplasma infections. The signs of eye lesions and a clinical case of an eleven-year-old girl diagnosed as having central recurrent multifocal exudative and hemorrhagic neurochoriorenitis of the right eye are given.

[Prevalence of toxoplasmosis and methods for its laboratory diagnosis].

The review presents data on the prevalence of toxoplasmosis in different countries and regions. It gives a comparative assessment of the methods of serodiagnosis of toxoplasmosis, which permit identification of specific immunoglobulins G, M, A, and E. The authors show a role of laboratory diagnostic techniques in the differentiation of the acute form of toxoplasmosis from its chronic form. They provide the currently available modifications of the basic procedure for enzyme immunoassay, molecular genetic methods, and a direct fluorescent antibody test for the detection of Toxoplasma gondii antigens. The current view of a cellular immune response and its role in monitoring the development of toxoplasmosis are considered.

[A gas chromatographic analysis of the fatty acid composition of Yersinia pestis]. / Gazokhromatograficheskii analiz zhirnokislotnogo sostava Yersinia pestis.

The fatty acid composition of Y. pestis strains, the causative agent of plaque, has been studied. Y. pestis cells have been found to contain great amounts of palmitoleic acid, methylenehexadecanoic acid, oleic acid with elaidic acid, palmitic acid and pentadecanoic acid. Lauric acid, myristic acid, 3-oxymyristic acid and methyleneoctadecanoic acid have been detected in moderate amounts. Vaccine strains, virulent museum and newly isolated strains, while differing in their antigenic structure, have proved to be uniform in their fatty acid composition. In Y. pestis cels, grown on a solid medium for a longer period and at higher temperature, an increased proportion of saturated acids is observed.

[The fatty acid composition of the cellular structures of Yersinia pestis]. / Sostav zhirnykh kislot letochnykh struktur chumnogo mikroba.

Fatty acid composition of lipopolysaccharides (LPS) and cytoplasmic membranes (CPM) of Yersinia pestis [correction of the plaque microbe] has been studied. Concentration of certain acids in the LPS content proved sharply different and thus the strains were separated into groups. High content of 3-oxymiristinic acid was a distinction of vaccine strains. Virulent strains of one of the groups regularly differed from the rest of virulent strains by high content of laurinic acid. Laurinic and two nonidentified acids, which were not found in the virulent cultures, were present in CMP lipids of vaccine strains.