Molecular typing of thermotolerant species of Campylobacter with ribosomal RNA gene patterns

Abstract

Ribosomal RNA gene restriction patterns (ribopatterns) of 72 strains representing Campylobacter jejuni (subspecies jejuni and doylei), C. coli, C. upsaliensis and C. lari including the urealytic (UPTC) biovar were determined using four common restriction endonucleases (HaeIII, HindIII, PstI and PvuII). The relative effectiveness of these enzymes for general molecular typing of the thermotolerant campylobacters was assessed. Ribotypes (HaeIII) were defined on the basis of computer-assisted numerical analysis. For C. jejuni, C. coli and C. lari, the HaeIII ribopatterns provided a high level of typability and discrimination, with patterns that were reproducible and easy to code for numerical analysis. There was evidence of diversity within three of the HaeIII types, and PstI ribopatterns proved the most reliable for detecting such differences. C. upsaliensis also could be ribotyped with HaeIII, but HindIII, PvuII and PstI were less satisfactory for this species. For such campylobacters, the HindIII ribopatterns generally were complex and difficult to compare, and the PvuII profiles provided the least discrimination. We conclude that the choice of restriction endonuclease is of critical importance when applying ribotyping to different species of Campylobacter. HaeIII ribopatterns were the most effective means of typing strains of different thermotolerant species of campylobacters and, when combined with PstI ribopatterns, offered a highly discriminatory basis for molecular typing.