Discriminatory power and application of ribotyping of Yersinia enterocolitica O:3 in an epidemiological study

Ribotyping performed with five restriction endonucleases was used in an attempt to subtype Yersinia enterocolitica serotype O:3 and also as a tool for clonal analysis. DNA from organisms under study (48 isolates from diarrheic human feces, 24 from food, and 5 reference strains) was tested by Southern hybridization using a DNA probe carrying an rRNA operon from Escherichia coli. Strains were grouped into seven ribotypes by the HindIII restriction endonuclease, into five ribotypes by Ncil, Bg/l, and Sa/l; and into two ribotypes by EcoRI, resulting in a discrimination index (DI) of 0.37, 0.17, 0.43, 0.13, and 0.03 for the five endonucleases. By combining the results obtained with two or more restriction endonucleases, a further discrimination was registered, the most efficient combination (in terms of discriminatory power vs. cost in work, time, and money) for routine typing being HindIII-Bg/l (9 types, DI=0.58). In the clonal analysis, results obtained with the five restriction endonucleases allowed us to define 11 groupings or clonal lines, which showed a remarkable degree of genetic heterogeneity (genetic distance coefficients between 0.03 and 0.73) and were grouped into two major clusters. One cluster included 93% of the strains and eight lines. At least two of the most frequent lines can be considered endemic in Asturias, Spain, because organisms belonging to these lines have been circulating and causing human yersiniosis in recent years and have also been isolated from commercial raw meat products. Two Ncil ribotypes from the series under study (92.2% of strains included in the prevalent cluster) were similar but not identical to ribotypes of 0.3 organisms from other geographic areas described in the literature, indicating that the genetic structure of prevalent human pathogens of this serotype is basically clonal.