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AIM: Analysis of clinical cases of tick-borne spotted fever (TSF) group rickettsiosis in 2005 - 2010. MATERIALS AND METHODS: General clinical, biochemical and serological parameters were determined in 10 tick-borne spotted fever group rickettsiosis patients who had visited various geographical regions of the World. RESULTS: TSF group rickettsiosis diagnostic criteria, optimal serological diagnostics timing were determined. Possible diagnostic errors, features of serological diagnostics and antibacterial therapy of this nosologic form are discussed. CONCLUSION: Indication for TSF examination are primarily epidemiologic including tick attachment indication and clinical data. Serological studies are positive only in 3 - 4 weeks after the onset of the infection and thus can not be used for early diagnostics.

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The technology of the purification of C.burnetii from the yolk-sac components of the cultivation medium, including differential centrifugation, membrane filtration, enzymatic hydrolysis, the standardization of the preparation thus obtained and its stabilization by the method of lyophilization, was developed. Antigens, prepared in accordance with this technology, were sufficiently active and highly specific. The possibility of their use in different serological tests (the complement fixation test and the indirect fluorescent analysis) was studied. In addition, these antigens were found to have good capacity for sorption on the solid-phase carrier (plates) and to be suitable for use in such highly sensitive test as the enzyme immunoassay.

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Species-specific antigenic erythrocyte diagnosticum (SAED) for the detection of antibodies to R.prowazekii was developed. The cells used as the basis for the preparation of the antigenic erythrocyte diagnosticum were formulated and tanned sheep red blood cells (FT-SRBC). Original indication antigens, solubilized with Triton X-100 from common R.prowazekii antigen for the complement fixation test, the antigen neutralization test and the fluorescent analysis, were used as sensitin. The optimum parameters for sensitizing FT-SRBC were established. The SAED thus obtained was stabilized by the method of lyophilization. In a separate series of experiments the specificity and activity of SAED were studied in comparison with those of preparations obtained with the use of different sensitins. The newly developed SAED was shown to be highly specific.

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The methods for obtaining antigenic and immunoglobulin preparations intended for the rapid serological diagnosis of coxiellosis were developed. The cells used as the basis for their preparation were formulated tanned sheep red blood cells. As sensitin, rabbit anti-C.burnetii immunoglobulin was used. The capacity for the detection of C.burnetii antigens in the passive hemagglutination test and specific antibodies in the passive agglutination test with the use of the our immunoglobulin erythrocytic diagnosticum was studied. To construct coxiellosis antigenic erythrocytic diagnosticum, the complex of membrane antigens obtained from C.burnetii strain M-44 was used as sensitin. The positive results of the approbatory trial of the erythrocytic diagnosticums made it possible to recommend both preparations for use in laboratory practice in the system of conjugated unidirectional serological reactions.

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For the first time the lyophilized fluorescent Fab-fragments of rabbit antibodies to horse serum protein, suitable for detecting different antigens and antibodies to these antigens, have been obtained by the specially developed method. The criteria to be used in the control of the antispecific fluorescent fragments of antibodies have been described and the methods of their control before and after lyophilization have been developed. The use of the antispecific fluorescent Fab-fragments of antibodies has been shown to considerably accelerate and simplify the indirect immunofluorescent assay.

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Parenteral administration of antiencephalytic gamma-globulin and of the Fab-fragments, extracted from it, to rabbits stimulated the formation of specific antibodies. The maximal blood serum specific antibody level in rabbits reimmunized with Fab-fragments was much lower than that following the reimmunization of rabbits with gamma-globulin. Antibodies specific to gamma-globulin and Fab-fragments were concentrated in the gamma-globulin fraction of the immune sera and could be referred to the immunoglobulins of G class.

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With the aid of the active anaphylactic reaction a comparative study was carried out of the reactogenic (anaphylactogenic) properties of gamma-globulin against tick-borne encephalitis from horse serum and of Fab-fragments isolated from it by gel-filtration. There proved to be statistically significant differences in the reactogenic properties of the preparations under study on condition of their separate use (reactogenic property of Fab-fragments was four times less than that of gamma-globulin). In case of a combined use of Fab-fragments and gamma-globulin, i.e. administration of the former after the sensitization of experimental animals with the latter, reactogenicity level of the two biopreparations proved to be practically the same.

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A comparative study was made of the serological properties and virus-neutralizing activity of antiencephalitis gamma-globulin and Fab-fragments isolated from it by gel-filtration. Horse immunoglobulins against the autumno-summer tick-borne encephalitis virus could be disintegrated with the aid of papaine to monovalent Fab-fragments which (according to the complement fixation reaction, the test of suppression of the complement fixation, and the HAIT) retained the serological activity whose level was compared with that of the serological activity of gamma-globulin. Fab-fragments possessed a marked virus-neutralizing activity. The mean value of a logarithm of the neutralization index was 2.65 +/- 0.2 for Fab-fragments and 3.74 +/- 0.38 for gamma-globulin (P less than 0.01).

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The authors present the results of immunization of horses-producers with a commercial antigen and the yolk culture of the living R. sibericus (strain K1) for the purpose of obtaining specific immune sera for many purposes. It was shown that the original combined scheme of immunization and reimmunization of horses, successfully approved in the preparation of immune sera to Rickettsia prowazeki also proved to be highly effective for obtaining the antisera to R. sibericus. Sera obtained after the primary immunization of horses could be successfully used as diagnostic sera, but they were of no use for the preparation of fluorescent antibodies and the highly active hemosensitin; as to the sera obtained after the remote reimmunization--they were universal raw material. Dry fluorescent antibodies to R. sibericus were prepared of the horse sera (their stain titre was 1 : 256--1 : 512) a highly active hemosensitin, and also dry commercial diagnostic sera for the IHT, CFR and the immunofluorescent reaction of microagglutination.

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