Burkholderia mallei genotyping based on different region analysis

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.