A method for differentiation of <Emphasis Type="Italic">Bacillus anthracis</Emphasis> strains and phylogenetically related species based on determination of the structural differences between chromosomal genes for biosynthesis of flagellin and methionine

Nucleotide sequence analysis of several genes responsible for definitive properties of the anthrax pathogen—motility and penicillinase activity—determined a chromosomal locus promising for interspecies differentiation. We demonstrated that the fliC gene encoding flagellin synthesis contains an extended region distinguishing B. anthracis strains from the majority of nonpathogenic and opportunistic bacilli. A novel method for anthrax pathogen indication and identification based on determination of the differences in the fliC and hom2 chromosomal genes structure has been proposed. A total of 60 strains of different Bacillus spp. (B. anthracis, B. cereus, B. thuringiensis, B. mycoides, B. megaterium, B. subtilis, etc.) were tested using two chromosomal DNA targets. The algorithm developed in this work permits detection of the pathogenic microorganism and reliably differentiation of it from other Bacillus spp. representatives. The introduction of primers complementary to specific sequences of pXO1 and pXO2 plasmids into multiplex PCR makes it possible to obtain additional information on the proposed virulence of the isolate.