A three-way ribotyping scheme for Salmonella serotype Typhimurium and its usefulness for phylogenetic and...

Ribotyping of Salmonella serotype Typhimurium strains was optimised as a tool for epidemiological and phylogenetic purposes. Of five restriction endonucleases evaluated on a series of 84 isolates, HincII, SalI and PvuII were the most useful, generating 13, 9 and 9 ribotypes with 17, 11 and 18 polymorphic restriction sites, and attaining a discrimination index (DI) of 0.81, 0.53 and 0.59, respectively. The combination of results from tests with the three enzymes provided further discrimination (19 ribotypes, DI=0.84). It proved useful for clonal analysis, defining 19 clonal lines with a remarkable degree of genetic heterogeneity, that were grouped into two major clusters (including 12 and 7 lines, respectively) at a significance level of 0.65. When the attributes of this system were compared with those of phage typing, it was found that ribotyping showed higher typability and sensitivity, supporting its use as an appropriate molecular method. In tracing the molecular epidemiology of Typhimurium strains in Asturias, six lines were found that could be considered endemic and were represented by organisms implicated in salmonellosis throughout the period of study; another four lines included organisms isolated from meat, water or both.