Restriction endonuclease analysis of nosocomial isolates of Clostridium difficile

A total of 110 clinical isolates of Clostridium difficile were analyzed by agarose gel electrophoresis by using both bacterial restriction endonuclease analysis (REA) and plasmid profiles. A total of 72 isolates were divided into 12 groups according to their REA patterns. Some 38 isolates exhibited unique patterns. Pattern A occurred in 20% of isolates. Isolates with patterns B, E, and G were cytotoxin negative. The remaining groups were cytotoxin positive. Multiple isolates obtained from two stool specimens were studied to examine the variation in REA profiles found in single specimens. In these specimens no variation in REA profiles was found. The stability of C. difficile was studied by examining sequential in vitro subcultures of a single isolate and strains isolated over a 4-month period from two long-term carriers. REA patterns were stable over time, both in vitro and in vivo. Because plasmid DNA was observed in 53% of isolates, plasmid profiles alone could not be used to study the spread of C. difficile; however, they were necessary for the interpretation of REA patterns in some instances.