[Role of laboratory methods in epidemic control of brucellosis outbreaks in Moscow zoo nursery].

Aim: Evaluation of the role of contemporary methods in epidemic control of brucellosis outbreaks among employees of auxiliary facilities in Moscow zoo nursery.

Materials And Methods: During 2003-2013, biannually, sera from more than 200 employees of the nursery, that work during periods of epizootics of small and large cattle in nursery auxiliary facilities, were studied. A complex of laboratory methods for brucellosis was used: variations of traditional serologic agglutination method in tubes--Wright's reaction (WR), on glass--Huddleston's reaction, Coombs' anti-globulin reaction; enzyme immunoassay with immune-dominant S-LPS; realtime polymerase chain reaction with primers based on genus target of BCSP31 protein gene.

[Molecular-genetic characterization of canine and rangiferine brucella isolates from different regions of Russia].

Goal: Comparative molecular-genetic characterization of Brucella isolates from dogs and reindeers in Russia by molecular-genetic typing methods.

Materials And Methods: 19 canine and 2 rangiferine Brucella isolates were studied by molecular typing methods based on PCR for differential species and biovar specific molecular targets and MLVA (multiple locus variable number tandem repeats analysis) using primers to 12 known variable loci.

Results: Using PCR for differential molecular targets, canine Brucella isolates were characterized as B.

[Use of multiple locus variable number tandem repeats analysis for the Brucella systematization].

Unlabelled: The methods of molecular-genetic differentiation to strain level acquire increasing significance in the current system of struggle with brucellosis. MLVA (multiple locus variable number tandem repeats analysis) was selected for molecular-genetic differentiation to strain level and simultaneous establishment of the genetic relationship of investigated Brucella strains. The goal of this work was MLVA typing of three pathogenic Brucella species strains with the analysis of stability of chosen loci, discrimination power and concordance to conventional phenotypic methods of the Brucella differentiation for use in systematization of brucellosis causing agents.

[Genetic characterization of the Brucella melitensis isolates from Mongolia, Russia, and Azerbaijan].

Unlabelled: The goal of this work was to provide comparative genetic characterization of the human and animal Brucella melitensis isolates from Mongolia, Russia and Azerbaijan using current molecular-genetic typing methods.

Materials And Methods: Twenty eight Mongolian (n = 18), Russian (n = 6), and Azerbaijan (n = 4) human and animal Brucella melitensis isolates were studied using 2 molecular typing methods based on PCR for differential species and biovar specific ORF (open reading frames) molecular targets and MLVA (multiple locus variable number tandem repeats analysis) using primers to 12 known loci.

Results: The PCR was used for differential molecular targets (all B.

Epidemiology chapter.

This chapter outlines the epidemiology of brucellosis in the Russian Federation and in five countries bordering Russia. Since the Soviet Union's dissolution, Russia and the newly formed independent republics have failed to maintain policies to control brucellosis and other zoonotic diseases. Many of these republics, due to weak animal control and prevention systems and dangerous food preparation practices, are still burdened with the human cost of brucellosis.

[Molecular genetics typing of Brucella circulating in several provinces of Mongolia].

Aim: Comparative molecular-genetic typing of Brucella strains isolated in Mongolia from different animal species as well as from humans.

Materials And Methods: Twenty-one strains of Brucella isolated from different hosts in 7 provinces of Mongolia were typed. Conventional phenotypic methods, genotyping by PCR with primers for genus- and species-specific differentiating targets of Brucella genes as well as multiple locus variable number tandem repeats analysis (MLVA) with 12 pairs of primers bounding locus variable tandem repeats of different length (from 134 bp to 8 bp).

Variable-number tandem repeat markers for identification of Brucella abortus 82 and 75/79-AV vaccine strains.

In Russia, live vaccine strains Brucella abortus 82 and B. abortus 75/79-AV have been widely and extensively utilized for specific prophylaxis of cattle brucellosis. To differentiate these vaccine strains from each other and laboratory collection of other vaccine (n=4), reference (n=15) and field Brucella strains (n=61), the multiple-locus variable-number tandem repeat (VNTR) analyses (MLVA) were used with 12 loci containing tandem repeats from 134bp to 8bp recently described for Brucella spp.

[Modeling of Brucella persistence in macrophage-like cells in vitro].

Aim: To develop model of chronic brucellosis infection in macrophage-like cells in vitro and to study properties of persistence of Brucella in them.

Materials And Methods: Infection of macrophage-like cells U937 and phagocytes B10.MLM with analysis of B.

[Highly sensitive enzyme-linked assay based on monoclonal antibodies for detection of Brucella antigens].

Mice monoclonal antibodies against lypopolysaccharides (LPS) of Brucella abortus has been obtained and characterized. The antibodies detected LPS of B. abortus, B.

[Molecular basis of Brucella persistence].

This review focuses on Brucella persistence. Data on Brucella--macrophage interaction and the role of molecular-genetic systems including homologues of Mos operon Rhizobium, a type IV secretion system, a flagellum apparatus and a "quorum sensing"--virulence factors using signal or effector molecules are updated. Brucella enters macrophages through lipid raft microdomains, avoids its bactericidal attacks, phagolysosome fusion, expressing a set of virulence genes and inhibits TNF-alpha secretion and apoptosis for persistence in macroorganisms.

[Enzyme immunoassay and polymerase chain reaction for evaluation of brucella persistence].

The complex approach, including the use of traditional bacteriological and serological methods, as well as the polymerase chain (PCR) reaction and the enzyme immunoassay (EIA), was used for evaluation of Brucella (the causative agents of brucellosis) persistence in the dynamics of the infectious process in patients with the acute and chronic forms of brucellosis as well as in experimentally infected laboratory animals. Sick humans and experimental animals were found to have positive PCR and EIA reactions at different periods of the disease. The use of these methods makes it possible to evaluate indirectly the persistence of Brucella.

[Molecular bases for brucella virulence].

The authors review published reports on the molecular bases of Brucella virulence, including type IV secretion proteins, S-lipopolysaccharide biosynthesis enzymes (O-antigen), regulatory proteins of various systems, and cellular metabolism proteins. High efficiency of modified transposon mutagenesis technique (selective labeled transposon mutagenesis) in search for virulence genes is shown. Analysis of DNA sequences of Brucella genome promotes identification of new virulence factors.

[Use of molecular-biological methods for identifying brucella in a comparative analysis of strains, isolated from sick dogs].

Ten strains isolated from sick dogs in 1998 in St. Petersburg were studied by traditional and molecular biological methods of Brucella identification. PCR study confirmed that the isolated cultures were Brucellae, and comparative study of the traditional phenotypical characteristics and protein and antigenic composition allowed referring all the isolated strains to B.

[Increased production of the protein antigen of Brucella melitensis in Escherichia coli K-12 cells].

The production of Brucella melitensis protein antigen with molecular weight of 38 kD in Escherichia coli K-12 cell lysates has been studied by immunoblotting with various antisera. E. coli strains differed by the vector plasmid and the size of B.

Development and evaluation of a rapid dipstick assay for serodiagnosis of acute human brucellosis.

A dipstick assay for the detection of brucella-specific immunoglobulin M antibodies was evaluated with 707 sera from 247 laboratory-confirmed brucellosis patients and 342 control sera from brucellosis-free individuals. These sera were collected from six different countries. The assay was found to be highly sensitive and specific.

[Sensitivity and adaptation of various Brucella species to acid shock].

The sensitivity to acid shock (pH 2.5, 2.8, 3.

[Use of Brucellar protein antigen synthesized in Escherichia coli cells in the enzyme immunoassay].

The possibility of using brucellar protein antigens with mol. weights of 18 and 38 kD, synthesized in E.coli cells, as sensitins on polystyrene plates in the enzyme immunoassay (EIA) for the detection of antibodies in the blood sera of patients with different forms of brucellosis.

[Comparative analysis of antigens from different Brucella species using immunoblotting with antisera from immunized rabbits].

Brucella antigens recognized by IgG antibodies in cell lysates from various Brucella species differing by the origin, biological, and virulent properties (including the reference, vaccine, and newly isolated strains) were compared by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins in SDS-cell lysates were separated by 12% SDS-PAGE and protein gels were stained with Coomassie brilliant blue R-250 and Silver reagent. SDS-PAGE showed differences in the protein profiles of 15 strains of different species.

[Comparative study of the antigenic composition of Brucella canis strains by immunoblotting].

Brucella cultures isolated from sick dogs have not been properly studied in Russia up to the present time. In 1994 a culture has been isolated from aborted fetus of a dog. Investigations by the traditional methods referred it to Brucella genus, species canis (strain K-01).

[An evaluation of the effectiveness of laboratory diagnostic methods for brucellosis].

The diagnostic value of bacteriological and serological methods for the laboratory diagnosis of brucellosis was studied. In the analysis of milk and cheese specimens Brucella cultures were isolated and differentiated as B.melitensis, biovar I, and B.

[The functional activity of the peripheral blood neutrophils in patients with chronic brucellosis].

The functional activity of peripheral blood neutrophils was studied by their main functional and metabolic parameters. The group of chronic brucellosis patients under study included 32 patients. The control group consisted of 35 practically healthy donors.

[Elaboration of a rapid method of determining the sensitivity of Brucella to antibiotics].

Optimal conditions were developed for determination of antibiotic sensitivity in Brucella by using enzyme immunoassay directly in the primary cultures of the material tested. The Brucella concentration in the material tested should be not lower than 1.10(6) microbial cells/ml and the time of culture incubation be 24 hours at 37 degrees C.

[Specific IgE antibodies in brucellosis].

Specific IgE antibodies and sensitization were assessed in patients with brucellosis in various forms. IgE antibodies were determined with enzyme immunoassay which demonstrated high specificity and sensitivity with conjugate against IgE antibodies in 509 serum samples. Specific IgE antibodies were present in most patients with acute, subacute or chronic brucellosis.