[Comparison of three Clostridium difficile culture media: interest of enhancing spore germination media?]

Abstract

Aim: Clostridium difficile is the most common agent of postantibiotic and nosocomial bacterial diarrhoea. Since the emergence of the highly virulent and epidemic strain NAP1/027 in Europe, it appears necessary to isolate C. difficile strains to realize an epidemiologic follow-up by molecular typing. The aim of this work was to compare three selective culture conditions for the isolation of C. difficile.

Methods: One hundred and thirty stools collected from patients hospitalized at Jean Verdier were swabbed on the commercial medium CLO (BioMérieux) and on a medium prepared at the laboratory (CCTa: Columbia, cefoxitine 8 mg/l, cycloserine 250 mg/l, horse blood 5 %, sodium taurocholate 0.1 %) with and without preliminary alcoholic shock (EtOH). C. difficile was isolated from 38 stools and colonies were counted on each medium.

Results: The fluorescence intensity of C. difficile colonies is comparable on CLO and CCTa-EtOH media, however their aspect is more characteristic on CLO. This medium appears very selective contrary to the CCTa medium on which an associated flora obstructs the fluorescence reading and requires a new isolation of the suspect strains. On average 30 times more colonies of C. difficile are counted on CCTa+/-EtOH than on CLO, suggesting the presence of great proportions of spores in the stools.

Conclusions: The medium CLO is successful for the isolation of C. difficile despite of its selectivity. Nevertheless, it appears interesting to associate a medium enhancing spore germination as the CCTa medium inoculated after alcoholic shock to increase the sensitivity of detection while being freed from conservation and transport conditions.