[The development of polymer immunoglobulin preparations to identify different serovars legionella pneumophilia in reaction of slide-agglutination].

The article deals with the results of study targeted to develop polymer diagnostic preparation to identify epidemically significant serogroups Legionella pneumophilia. The preparation combines rate of record (1-5 min) of reaction of paragglutinining preparations with color visualization and demonstrative of reaction of volume agglomeration with polymer diagnosticums. The specially synthesized polymer microspheres were sensibilized with serums enriched with antibodies to lipopolysaccharide of corresponding serovar L. pneumophilia. The derived immunoglobulin diagnostic preparations detect agent of legionellesis in the reaction of slide-agglutination on glass during 1-5 min. The polymer diagnostic preparations provide positive reaction with culture of corresponding serovar and no reaction with other gomologic and geterologic agents of infectious diseases.

[Comparative study of the biological properties of various nutrient media used for the isolation of Legionella pneumophila].

Three diagnostic selective media used for the isolation of Legionella pneumophila were compared. These included BCYEAalpha (Oxoid, a reference medium), BCYEAalpha (Hi Media), and elective legionellosis medium (ELM) developed at the Rostov-on-Don Research Institute for Plague Control. The virulent L. pneumophila strain Philadelphia-1 (LD50 was 10(5) CFU for guinea-pigs) was used a test culture. The susceptibility of the media was determined, by culturing 10(-6) and 10(-7) dilutions of the suspension of the macerated Legionella-infected guinea-pig spleen, as well as the suspension of a culture of this strain (100 CFU) and 6 L. pneumophila cultures freshly isolated from water. The BCYEAalpha (Oxoid) and ELM media demonstrated the similar growth characteristics (chi2 < 0.7; p = 0.05) while the BCYEAalpha (Hi Media) medium showed a low susceptibility. The BCYEAalpha (Oxoid) and ELM media were first found to be successful in detecting Legionella in viable, but nonculturable state, induced by the following factors: 1) starvation in distilled water; 2) exposure to hydroquinone (oxidative shock, and 3) elevated temperature (56 degrees C). The BCYEAalpha (Oxoid) and EML media did not differ either in their ability to suppress extraneous microflora and to maintain stable initial pH under the conditions of incubation of culture plates, as well as in their Na+ concentration (15-19 mmol/l). However, the BCYEAalpha (Oxoid) medium exceeded the ELM one in the growth rate and diameter of Legionella colonies. Two L. pneumophila cultures were isolated in the BCYEAalpha (Oxoid) and EML media used in the field experiment studying 15 water samples from different hot water supply systems. Thus, the conclusion can be drawn that the EML medium is comparable with the reference BCYEAalpha (Oxoid) medium in its susceptibility and ability to detect Legionella in both vegetative and viable, but nonculturable states and is suitable for practical application.

[Novel diagnostic kit on the basis of stained polymeric carriers for epidemiologic surveillance in natural foci of Crimean-Congo hemorrhagic fever].

AIM: To study diagnostic value of developed antigenic polymeric diagnostic kit for epidemiologic surveillance in natural foci of Crimean-Congo hemorrhagic fever (CCHF) in South federal district. MATERIALS AND METHODS: Novel antigenic diagnostic kit on the basis of polymeric microspheres for reaction of volume agglomeration was developed. The kit is designed for detection of virus-specific antibodies in human serum and in serum of agricultural animals. RESULTS: Laboratory and field trials of the kit showed its high diagnostic potency, it was included in methodical recommendations "Organization and accomplishment of measures against Crimean-Congo hemorrhagic fever on the territory of its natural foci in Russia". Use of antigenic polymeric kit for epidemiological surveillance allows for more complete and systemic understanding of CCHF epidemic process. CONCLUSION: At present, the diagnostic kit is successfully used, alongside with ELISA and PCR, on different levels of epidemiologic surveillance for CCHF in Rostov region.

[Study of Legionella pneumophila strains isolated from environmental objects].

The paper presents the results of studying the biological properties of Legionella pneumophila strains isolated from environmental objects. Elective legionellosis medium (ELM) has been found to be suitable for the isolation of the causative agent from the starting material and to be as sensitive as CYE (Oxoid company) containing growth and selective additives. Polymerase chain reaction (PCR) with a home-produced commercial test system used to detect L. pneumophila DNA enables identification of the causative agent, including its species. Hyperimmune sera against L. pneumophila 1-7 serogroups used in slide-agglutination and agglutination, as well as a series of co-agglutinating diagnosticums for legionellosis 1-7 serogroups make it possible to identify even the serogroups of L. pneumophilla. Comparative analysis of the virulence of L. pneumophila cultures in vivo and in vitro allows recommendation that practical laboratories should employ a simple NaCl resistance test, which can be used as a guide virulence test.

[Serologic diagnostics of Legionella infection].

Technical approaches to construction of preparations for serologic diagnostics of Legionella infection were presented in the article; antigenic- and immunoglobulin-based diagnostic kits with known characteristics were developed. Immunogenic properties of protein and lypopolysaccharide antigens, which have diagnostic value, were studied; similarity of protein antigens from 7 serogroups of L. pneumophila was demonstrated. Soluble antigen with known composition was obtained and used for the development of antigen-based polymeric kit for diagnostics of Legionella infection. On the basis of hyperimmune sera, immunoglobulin-based polymeric diagnostic kit and array of coagglutinating diagnostic kits for the mentioned 7 serogroups were developed. Antigen-based polymeric diagnostic kit was recommended for licensure.

[Detection of lipase in Vibrio cholerae serogroup O1 in reaction of volumetric agglomeration].

Study showed that El-Tor strains of V. cholerae isolated from different sources produce lipase for hemolysis after cultivation during 24 h on meat-peptone broth independently from their toxigenic and hemolytic abilities. Study of 3- and 4-hours broth cultures of vibrios revealed possibility to differentiate between hemolytic nontoxigenic strains and toxigenic nonhemolytic ones. Using antilipaze diagnostic kit it was possible to differentiate El-Tor vibrios from vibrios of classic biovar basing on lipase production 24 h after cultivation on meat-peptone broth that was evident in El-Tor vibrios but not in classic biovar strains.

[Development of antilipase immunoglobulin polymer diagnosticum for the detection of Vibrio cholerae eltor, possessing hemolytic and lipase activity].

The possibility of using a heterogeneous, but structurally similar antigen--the commercial preparation of Pseudomonas sp. lipase (Sigma, USA)--for the development of polymer diagnosticum aimed at determination of lipase production in cholera vibrios was shown. The new diagnosticum (antilipase antibodies) on a polymer carrier was used in the serological volume agglomeration test for the detection of hemolytic atoxigenic V. eltor, obtained from environmental, objects, which produced lipase in 80% of cases. The differentiating capacity of the diagnosticum was confirmed on 120 V. eltor cultures isolated from environmental objects. The newly developed diagnosticum makes it possible to determine the lipase activity in cholera vibrios of different biovars and serovars.

[Epidemiologic characterization of Brucellosis under modern conditions]. / Epidemiologicheskaia kharakteristika brutseleza v sovremennykh usloviiakh.

The spread of brucella infection under modern conditions of cattle breeding were studied with the foci in the Rostov region taken as an example. The main epidemiological features and tendencies of epidemiological importance were established. To find out specific antigen in different environmental objects, the approbation of the enzyme immunoassay test systems was carried out and the possibility of their use at the institutions of the State Sanitary and Epidemiological inspection was shown.

[The characteristics of cholera prevalence and of the performance of epidemic-control measures in small settlements of Dagestan]. / Osobennosti rasprostraneniia kholery i provedeniia protivoépidemicheskikh meropriiatii v nebol'shikh naselennykh punktakh Dagestana.

The present work deals with summarizing the experience obtained by the specialized antiepidemic brigade of the Rostov-on-Don Research Institute for Plague Control in the work on the liquidation of cholera in some regions of Daghestan with a view to discussing the problems of improvement of anticholera measures. The characteristic features of the epidemic process were its explosive character, sparseness of the foci of infection, the prevalence of its transmission through everyday contacts (family contacts and intensive tribal contacts) and essential delays in taking anticholera measures due to sudden appearance of outbreaks, remoteness of small settlements and the lack of manpower and means for carrying out anticholera measures at a given place and time, as well as delays in epidemiological analysis carried out by local health service bodies. Delays in carrying out such measures led to the spread of infection both within settlements and in the whole region and further in the republic. The epidemic process was complicated by the antibiotic resistance of V.cholerae strains circulating on this territory. All these factors formed specific epidemic situation which introduced amendments into the organization of anticholera measures.

[The characteristics of the antibacterial therapy for cholera in Dagestan]. / Osobennosti antibakterial'noi terapii kholery v Dagestane.

Wide circulation of antibiotic-resistant Vibrio cholerae strains again gives prominence to the problem of etiotropic therapy. The results of the treatment of 428 persons infected with V.cholerae (237 cholera patients and 191 Vibrio carriers) in different regions of Daghestan during the outbreak of epidemic in 1994 are presented. The main criterion of the effectiveness of antibacterial therapy was the determination of the percentage of bacterial relapses. The sensitivity of 118 V.cholerae strains to different antibacterial preparations was studied by the method of serial dilutions. After the clinical use of chloramphenicol 29.7% of bacterial relapses were registered, the in vitro resistance of V. cholerae being 32-64 mkg/ml. After the use of tetracycline 16.5% bacterial relapses were registered with in vitro resistance being the same. The use of the combination of these preparations gave 15% of bacterial relapses. Furazolidone gave 4.3% of bacteria relapses, while after the use of ciprofloxacin 2.8% of bacterial relapses were registered with in vitro sensitivity equal to 0.25-0.5 mkg/ml. Ciprofloxacin was recommended for the treatment of cholera patients and furazolidone, for the treatment of Vibrio carriers.

[The preservation of the causative agent of cholera in the water supplies of the central regions of Dagestan (experimental data)]. / O sokhranenii vozbuditelia kholery v vodnykh istochnikakh tsentral'nykh raionov Dagestana (éksperimental'nye dannye).

The acidic pH of water of surface water reservoirs in Izberbash and two adjoining regions, including sea water, seems to be unfavorable for the prolonged preservation of Vibrio cholerae eltor, but additional ecological investigations are necessary to study the possibility for infection to take root at this territory. Water from the Zam-Zam spring, if contaminated with V. cholerae, may serve as a transmission factor, but the duration of its action is limited by the survival term of V. cholerae. The water route of transmission did not play any essential role in the spread of cholera in the central regions of Daghestan.

[The complex study of the epidemic situation and of the immunological structure of the population in brucellosis foci]. / Kompleksnoe izuchenie épidemicheskoi situatsii i immunologicheskoi struktury naseleniia v ochagakh brutselleza.

As the result of the complex clinico-laboratory examination of 265 cattle breeders working on 5 farms where cases of brucellosis among agricultural animals had been detected, 62 persons (21.8%) were found to have the positive reaction to brucellosis. A strikingly high proportion (38.3%) of infected persons was found among the workers of a cattle-feeding complex. Out of 62 persons with the positive reaction to brucellosis, 32 had different clinical manifestations of the disease. Treatment for all persons who needed it was organized, the rest were placed under dispensary observation. Regular complex clinico-laboratory examinations of cattle breeders to brucellosis were recommended.