Flagellin gene sequence evolution in Salmonella.

Salmonella exhibits 70 serologically distinct flagellins, used internationally to diagnose and track infections. The terminal sequences of flagellin protein subunits are conserved in a range of bacteria and are here used as evolutionary markers to reveal how new serotypes arise. Terminal sequences of flagellins that exhibit factors g or m (G-group) were distinct from other Salmonella antigens (Non-G-group) and cluster more closely with Escherichia coli. It is postulated that G-group flagellins were inherited from a common ancestor of E. coli and Salmonella and that these antigens were among the original set in Salmonella. Sequence differences at the 5' termini may prevent recombination between co-infecting strains. Evidence of increased variation of flagellin in rare biphasic G-group serotypes suggests that the presence of a second flagellin locus allows mutation of the G-group flagellin. FljB probably arose from a single duplication of a Non-G gene, since which synonymous mutations resulted in the fljB-specific sequence at the 5' termini.

Towards the development of a DNA-sequence based approach to serotyping of Salmonella enterica.

BACKGROUND: The fliC and fljB genes in Salmonella code for the phase 1 (H1) and phase 2 (H2) flagellin respectively, the rfb cluster encodes the majority of enzymes for polysaccharide (O) antigen biosynthesis, together they determine the antigenic profile by which Salmonella are identified. Sequencing and characterisation of fliC was performed in the development of a molecular serotyping technique. RESULTS: FliC sequencing of 106 strains revealed two groups; the g-complex included those exhibiting "g" or "m,t" antigenic factors, and the non-g strains which formed a second more diverse group. Variation in fliC was characterised and sero-specific motifs identified. Furthermore, it was possible to identify differences in certain H antigens that are not detected by traditional serotyping. A rapid short sequencing assay was developed to target serotype-specific sequence motifs in fliC. The assay was evaluated for identification of H1 antigens with a panel of 55 strains. CONCLUSION: FliC sequences were obtained for more than 100 strains comprising 29 different H1 alleles. Unique pyrosequencing profiles corresponding to the H1 component of the serotype were generated reproducibly for the 23 alleles represented in the evaluation panel. Short read sequence assays can now be used to identify fliC alleles in approximately 97% of the 50 medically most important Salmonella in England and Wales. Capability for high throughput testing and automation give these assays considerable advantages over traditional methods.