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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.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.A number of methods for identification and typing of glanders and melioidosis causative agents is proposed.

The development of methods for glanders agent genotyping is urgent due to the high pathogenicity of this microbe, lack of effective preventive measures, and threat of the use of Burkholderia mallei as a biological weapon. In this study, we propose a scheme for genotyping of B. mallei strains based on different region analysis (DFR). The choice of variable loci specific for different strains of the glanders agent was performed by analyzing the annotated whole-genome sequences of the B. mallei strains. Primers and fluorescence probes were designed for nine selected loci. The amplification conditions for different regions were optimized in two variants: with electrophoretic detection and hybridization-fluorescence detection in the strip format. DFR analysis was assessed in 14 B. mallei strains. The strain genetic profiles revealed that developed DFR-typing was characterized by a high discrimination power (Hunter–Gaston index was 0.92), reproducibility, rapidity, easy interpretation, and applicability for epidemiological surveillance of glanders.

Development of the genotyping methods of glanders agent is urgent due to its high pathogenicity, lack of effective preventive measures and threat of the use of Burkholderia mallei as a biological weapon. In this work we proposed a scheme for the typing of the B. mallei strains based on different region analysis (DFR). The choice of variable loci differentially presented in various strains of glanders agents was performed by analyzing annotated whole-genome sequences of the B. mallei strains. Primers and fluorescence probes were designed for 9 selected loci. The amplification conditions for different regions were optimized in two variants: with electrophoretic detection and hybridization-fluorescence detection in the strip format. The possibility of applying the DFR analysis to genetic characterization of strains was assessed in 14 B. mallei strains. The genetic profiles of the studied B. mallei strains revealed that the developed DFR-typing scheme was characterized by high discrimination power (Hunter-Gaston index value was 0.92), reproducibility, rapidity, easy interpretation, and applicability for epidemiological surveillance of glanders.

The glanders pathogen (Burkholderia mallei) is a highly pathogenic microorganism for humans and animals. In this article we studied the possibility of using genotyping methods for the differentiation of B. mallei strains. We characterized 14 strains of the glanders pathogen using amplification with random primers (randomly amplified polymorphic DNA, RAPD) and multilocus sequence typing (MLST). It was shown that RAPD was the best of the methods used for differentiating B. mallei strains, and it may therefore be more useful for molecular-epidemiological investigations.

Burkholderia mallei is highly pathogenic microorganism for both humans and animals. In this work, the possibility of the use of the genotyping method for differentiation between strains of B. mallei was studied. A collection of 14 isolates of B. mallei was characterized using randomly amplified polymorphic DNA (RAPD) and multilocus sequence typing (MLST). RAPD was the best method used for detecting strain differences of B. mallei. It was suggested that this method would be an increasingly useful molecular epidemiological tool.