Bacteriophage and bacteriocin typing scheme for Clostridium difficile

Abstract

The study of the epidemiology of infection with Clostridium difficile would be aided by a way to type individual bacterial isolates. We therefore sought bacteriophages for use in typing. With mitomycin C exposure (3 micrograms/ml), filtrates from 10 strains of C. difficile had plaque-forming lytic activity on other C. difficile strains. Individual phage were passaged and made into high-titer stock preparations for typing. Electron microscopy revealed tailed phage particles from one such preparation. In addition to phage, inhibitory activity without distinct plaque formation consistent with bacteriocins was observed for 20 strains. C. difficile isolates from 16 patients taken 1 to 14 days apart were similar in their phage sensitivity pattern, whereas isolates from separate geographic locations showed a great variety of patterns. We conclude that bacteriophage should be useful for typing strains of C. difficile.

Recent advances in the genetics of the clostridia

“…Disruption of industrial scale solventogenic fermentations by bacteriophages has also been documented (see [3]). Finally, bacteriophages have been used for typing strains of C. difficile [50].…”

Development of a new PCR-ribotyping method for Clostridium difficile based on ribosomal RNA gene sequencing

“…This bacterium is considered to be the main agent of nosocomially acquired diarrhea among adults [2]. Many various typing methods have been developed to investigate nosocomial outbreaks of C. di¤cile, some based on phenotype (lysotyping [3], serogroup-ing [4], SDS-PAGE [5], immunoblotting [6]) and others on genotype (plasmids [7], REA [8], ribotyping [9], PFGE [10], RAPD [11]). Recently, Kostman et al developed a method of PCR-ribotyping that was shown to be reproducible, easy to perform and cost-e¡ective [12].…”

Immunoblotting to demonstrate antigenic and immunogenic differences among nine standard strains of Clostridium difficile

“…Microbiol. 20:604, 1984) with the use of the bacteriophage method for typing described by Sell et al (8). Poxton et al (6) used immunoblotting with antiserum raised against one C. difficile strain to demonstrate that a single strain was involved in a hospital outbreak in Sweden.…”