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Biomedical subjects

D Linton

Publications and source records attributed to D Linton.

6 recordsLinked to original sources

Campylobacter helveticus sp. nov., a new thermophilic species from domestic animals: characterization, and cloning of a species-specific DNA probe.

An atypical group of thermophilic catalase-negative Campylobacter strains, the 'CH' (Swiss) group, can be recovered from faeces of domestic cats and dogs after selection by filtration, or with the antibiotic cefoperazone. This group of strains shows no relative DNA homology with any species in rRNA superfamily VI (Vandamme et al., 1991, International Journal of Systematic Bacteriology 41, 88-103) except with four thermophilic Campylobacter species, notably C. upsaliensis. The group is homogeneous and possesses a DNA base composition, cellular morphology at the electron microscope level and phenotypic properties characteristic of Campylobacter. Nonetheless it is distinct from known species of Campylobacter in terms of conventional bacteriological tests, total cellular protein profile, rRNA gene profile, and genomic DNA homology. On the basis of an integrated study of phenotype and genotype, we conclude that these bacteria constitute a previously undescribed species for which we propose the name Campylobacter helveticus sp. nov. A species-specific recombinant DNA probe was cloned from the designated type strain (NCTC 12470) for use in identification and further analysis of the epidemiology, pathogenicity and transmission of C. helveticus.

Animals

16S rrn gene copy number in Helicobacter pylori and its application to molecular typing.

The copy number of the genes encoding 16S ribosomal RNA was analysed for the genomes of geographically diverse strains of Helicobacter pylori, and restriction site variation within and around the genes was characterized. A DNA probe of 550 bp was amplified by the polymerase chain reaction from genomic DNA of the type strain NCTC 11637. This probe constituted a sequence internal to the 3' end of the 16S rrn gene. Homology profiles were compared for genomic Southern blots made with four restriction enzymes cutting within and outside the probe sequence. A copy number of two was established for all 12 strains analysed. This approach yielded significantly simpler data than does conventional 'ribotyping ' of H. pylori. It was equally discriminatory, however, and provided strain-specific 16S rrn gene 'signatures'. These represent both fundamental physical-genetic information and a novel approach to typing this gastric pathogen.

Bacterial Typing Techniques

Ribosomal RNA gene restriction patterns of Helicobacter pylori: analysis and appraisal of Hae III digests as a molecular typing system.

Ribosomal RNA gene restriction patterns (ribopatterns) of 162 strains of Helicobacter pylori from 93 patients were studied to assess their suitability for use as the basis of a molecular typing system. Computer-assisted numerical analysis of Hae III ribopatterns of 122 strains from 9 countries in 4 continents showed that almost every strain had a distinct and unique ribopattern and only strains from the same individual were genomically matched in all bands or with minor (1-2 band) differences. Hae III ribopatterns offered high typability and reproducibility but were too discriminatory for large-scale epidemiological typing purposes because no rational basis for grouping strains was evident. In contrast, composite band profiles based on 21 band loci within the Hae III ribopatterns, which were used to compare strain sets selected on toxigenicity and geographical origin, were more conserved. Minor differences between some strain sets in several band loci were detected but generally the composite profiles were a reproducible feature of H. pylori. We conclude that Hae III ribopatterns provide an excellent fingerprint for small-scale studies of genomic variation in defined populations, such as sequential patient isolates, but are too specific for general typing of H. pylori.

Blotting, Southern