[Studying the possibility of using immuno-enzy test system for evaluating anti-pertussis immunity.]

A research objective - to study the possibility of using the ELISA Anti-K enzyme immunoassay system to evaluate anti-pertussis immunity. А comparative assessment of the content of co-crank antibodies in the blood serum of adults, pregnant women and children 6 years old in the agglutination test, in the test system "Anti-K ELISA" and test systems of foreign production was carried out. The "Anti-K" IFA test system makes it possible to detect the level of specific antibodies to both the whole cell and cell-free pertussis component of the vaccine at any stage of the vaccination cycle. This diagnostic test can be used to determine the tactics of immunization, and to assess population immunity.

[SAFETY AND IMMUNOGENICITY OF A NATIONAL COMBINED VACCINE AGAINST PERTUSSIS, DIPHTHERIA, TETANUS, HEPATITIS B AND Hib-INFECTION, CONTAINING ACELLULAR PERTUSSIS COMPONENT, DURING IMMUNIZATION OF ADULTS].

AIM: Study safety, reactogenicity and immunologic effectiveness of a national combined vaccine against diphtheria, pertussis (acellular component), tetanus, hepatitis B and Hib-infection during immunization of volunteers aged 18-60 years. MATERIALS AND METHODS: The study was carried out in accordance with ethical standards and requirements, regulated by Helsinki declaration and Good clinical practice (ICHGCP). In a simple non-randomized clinical trial 20 adult volunteers took part, the mean age of those was 46.9 years. RESULTS: Registered: post-vaccination reactions (both local and systemic) were mild and of moderate degree of severity, stopped independently after 2-3 days without administration of drug treatment. Postvaccinal complications were not noted. Parameters of general and biochemical analysis of blood, urine, IgE content in dynamics of immunization were within normal limits. A single administration of aAPDT--HepB+Hib to individuals aged 18-60 years resulted in development of antibodies against all the components of the preparation. Seroconversion factor fluctuated from 6.9 to 53.5: CONCLUSION: The results obtained allow to recommend the vaccine for evaluation of its safety, reactogenicity, immunologic and prophylaxis effectiveness in randomized clinical observation trials in children.

[Epidemiologic diagnostic of nosocomial suppurative-septic infections of Pseudomonas etiology based on intraspecies typing of causative agent].

AIM: Scientific justification of optimization of epidemiologic diagnostic of suppurative-septic infection (SSI) caused by Pseudomonas aeruginosa based on comparability of antibiotic sensitivity and beta-lactamase production. MATERIALS AND METHODS: Intraspecies typing of 37 P. aeruginosa strains isolated during microbiological monitoring of 106 patients and 131 objects of clinical environment of surgical and obstetrician hospitals by using a complex ofphenotypic and molecular-biological methods including determination of sensitivity to antibiotics by serial dilutions method and PCR-diagnostics with determination of TEM, SHV, CTX, OXA, MBL, VIM genes was performed. RESULTS: P. aeruginosa strains combined into groups by isolation location during studies turned out to be heterogeneous by sensitivity to antibiotics and beta-lactamase production that allowed to form subgroups of strains by focality attribute. Isolates recovered from different SSI foci had significant differences in minimal inhibitory concentration (MIC) reaching 1024 times. MIC parameter within subgroups did not exceed 8 - 16 consequent dilutions. CONCLUSION: Use of a complex of phenotypic and molecular-biologic methods of causative agent typing including determination of sensitivity to antibiotics by serial dilutions method and evaluation of beta-lactamase production allowed to establish a mechanism of development of SSI epidemic process caused by P. aeruginosa, detect origins and reservoirs of infection in hospital, modes and factors of transmission and reach maximum justification of epidemiologic control and prophylaxis measures of localization of foci of nosocomial infections of pseudomonas etiology.

[Risk factors of development of nosocomial pyogenic and septic infections in maternity hospitals].

During prospective epidemiological surveillance cases of pyogenic and septic infections (PSI) in mothers and newborns in two maternity hospitals were studied using standard case definition and leading risk factors of their development were revealed. These factors differed in two hospitals and were connected mainly with high level of patients colonization, contamination of the environment by nosocomial strains of microorganisms, and degree of participation of mother's relatives in delivery. It was shown that permission to relatives for presence on delivery did not influence on the rate of PSI. Specificity of risk factors of PSI in mothers and newborns dictates necessity to determine them in each maternity hospital.

[Evaluation of the reactogenicity and immunogenic potency of combined vaccine "Bubo-Kok" in the immunization of children against diphtheria, tetanus, pertussis and viral hepatitis B]. / Otsenka reaktogennosti i immunogennosti kombinirovannoi vaktsiny "Bubo-Kok" pri immunizatsii detei protiv difterii, stolbniaka, kokliusha i virusnogo gepatita B.

Combined vaccine "Bubo-Kok" is characterized by safety and high immunological activity. The number of postvaccinal reactions in children aged 1 and 2 years, immunized with vaccine "Bubo-Kok", was not statistically different from those in groups of children immunized with adsorbed DPT vaccine, as well with such vaccine in combination with vaccine against hepatitis B. After the completion of the primary course of immunization 100% of children had protective antibody titers against diphtheria, tetanus and hepatitis B. Antibody titers against pertussis, equal to or exceeding protective titers, were found in more than 70% of immunized children. The immunogenic potency of vaccine "Bubo-Kok" with respect to all its components was not inferior to that of adsorbed DPT vaccine and vaccine against hepatitis B, when introduced simultaneously in different areas of the body. Vaccine "Bubo-Kok" successfully passed state trials and was recommended for registration.

[Reactions to and immunogenic properties of the combined vaccine Bubo-M for immunization of children against diphtheria, tetanus and viral hepatitis B]. / Reaktogennost' i immunogennost' kombinirovannoi vaktsiny Bubo-M pri immunizatsii detei protiv difterii, stolbniaka i virusnogo gepatita B.

Bubo-M, the first Russian combined vaccine, was found to have low reactogenicity. The difference between the number of postvaccinal reactions in the group of children immunized with Bubo-M (25.9%) and those in the group of children who had been simultaneously injected into different sites of the body with ADS-M toxoid (adsorbed DT toxoid with reduced antigen content) and hepatitis B vaccine (26.7%) was not statistically significant. Following immunization a considerable increase in the level of diphtheria and tetanus antibodies (p < 0.005) was observed in all children (100%), the level of HBs antibodies in the group of children immunized with Bubo-M (the geometric mean titer: 13,721 IU/l) essentially exceeding that observed in the control group injected with ADS-M toxoid and hepatitis B vaccine (the geometric mean of the titer: 2,441 IU/l). Bubo-M was duly registered and allowed for industrial production and medical use.

[Evaluation of reactogenicity and immunogenicity of Bubo-M, the Russian combined vaccine for immunization of adults against diphtheria, tetanus and viral hepatitis B]. / Otsenka reaktogennosti i immunogennosti otechestvennoi kombinirovannoi vaktsiny Bubo-M pri immunizatsii vzroslykh protiv difterii, stolbniaka i virusnogo gepatita B.

Bubo-M, the first Russian associated vaccine, was found to have low reactogenicity and high immunogenic potency. The frequency of postvaccinal reactions in the group of persons immunized with Bubo-M (20%) appeared to be considerably lower than among persons who received the combined injection of adsorbed DT toxoid with reduced antigen content and vaccine against hepatitis B (47.7%). Following the course of vaccination the level of anti-HBs considerably exceeded the protective level. Immune response to the diphtheria and tetanus components of Bubo-M exceeded that observed after immunization with absorbed DT toxoid with reduced antigen content (p < 0.05).

[The assessment of the parallelism in the results of measuring an analyzable substance and the calibrator in immunological assays]. / Ob otsenke parallelizma rezul'tatov izmereniia analiziruemogo veshchestva i kalibrovatelia pri immunologicheskikh ispytaniiakh.

The method for checking the results of measurements of the analyzed substance and the calibrator for parallelism the immunological testing kit ABICAP-test-DIPHTHERIA, produced by ABION (Germany), is described. The international standard sample of diphtheria antitoxin human serum was used as calibrator, and human serum No. 18, obtained from the Biomed Group (Russia), was used as the substance to be analyzed. The described method made it possible to detect the parallelism of "dose-response" curves with the relative error not exceeding 20%. Such accuracy proved to be sufficient for practical purposes and was in agreement with the amplitude of oscillations in measured optical densities. The shift of graphs along the abscissa made it possible to estimate the concentration of the serum as exceeding the concentration of the standard sample 2.3 times. The use of analytical approximation for the standard seems to be an important feature in the described method.

[Use of membrane separation methods in the production of Clostridium perfringens toxoid]. / Ispol'zovanie membrannykh metodov razdeleniia v izgotovlenii anatoksina Clostridium perfringens.

The dynamics of ultrafiltration was found to depend on the composition of the nutrient medium in which C.perfringens toxin accumulated. A new composition of casein-pancreatic medium with the reduced content of high-molecular peptides ensuring a high effect of membrane separation was developed. The influence of the main factors (pressure, concentration, the number of diafiltration cycles) on the characteristics of the membrane separation of type A C.perfringens culture. The optimum conditions for the purification of C.perfringens toxin by ultrafiltration were established with due regard to the data obtained in this investigation. Toxin concentrates were found to contain a fraction inhibiting their sorption capacity, and the method of the elimination of the inhibiting component was developed. C.perfringens toxoids, considerably exceeding commercial preparations in specific activity and immunogenic potency, were obtained.

[Certification of a kit for immunologic tests]. / O sertifikatsii nabora dlia immunologicheskikh ispytanii.

A diagnostic kit for the determination of IgG antibodies to diphtheria toxin ("Abion", Germany) the use of analytical columns "Abicap" was studied in the specialized laboratory of the Tarasevich State Control Institute with a view to determine the possibility of its official registration in the Russian Federation. The kit was evaluated by the following criteria: the correspondence of the specified properties of the preparation to those actually observed; meeting the norms stated in the documents supplied with the kit and WHO requirements; the presence of advantages in comparison with the enzyme immunoassay. In the course of the trial positive results were obtained on all these points.

[Detection of exotoxins of the pathogens of anaerobic gas infection using enzyme immunoassay]. / Vyiavlenie ékzotoksinov osnovnykh vozbuditelei anaérobnoi gazovoi infektsii s pomoshch'iu immunofermentnogo analiza.

To detect the exotoxins of the causative agents of the main anaerobic gas infection (Clostridium perfringens, C.oedematiens, C.histolyticum, C.septicum) a rapid and easily reproducible variant of the enzyme-linked immunosorbent assay (ELISA), based on the use of the sandwich modification activated due to avidin-biotin interaction, was proposed. The possibility of using the avidin-biotin variant of ELISA for the detection of C.perfringens, C.oedematiens, C.histolyticum and C.septicum toxins in experimental gas gangrene in guinea pigs was shown. The method made it possible to reproducibly detect 0.02-0.2 ng of antigens (gangrene toxins-toxoids) with the immunoassay being highly specific.

[A multistage metrologic approach to the problem of evaluation of the results of the quantitative enzyme immunoassay determination of antitoxic antibodies]. / O mnogoétapnom metrologicheskom podkhode k probleme otsenke rezul'tatov kolichestvennogo immunofermentnogo opredeleniia antitoksicheskikh antitel.

To evaluate a kit for the enzyme immunoassay (ELISA), the metrological approach was used: the total error of the method for the quantitative determination of antibodies, regarded as a multistage process, was determined as the result of the accumulation of errors made in measurements at different stages. The proposed algorithm made it possible to attest the positive control serum, to determine the total error of measurements, to mark a linear section in a limited range of values on a graduation diagram. This led to obtaining well-grounded results, comparable with the results of the reaction of neutralization at good correlation (lc = 0.9).