USE OF GENE AMPLIFICATION TO DETECT CLOSTRIDIUM-DIFFICILE IN CLINICALSPECIMENS

The combined use of an enrichment broth and gene amplification followi ng simple DNA extraction to detect toxigenic strains of Clostridium di fficile in feces was investigated by examining feces from 329 cases of suspected C. difficile infection. DNA was extracted by heating the wa shed centrifuged deposit from the broth in a microwave oven. For compa rison, specimens were tested concurrently using standard methods for c ulture and cytotoxin testing. Amplified fragments were identified by m olecular weight estimation, restriction enzyme digestion patterns and Southern blot hybridization. The combination of an enrichment broth fo llowed by gene amplification was shown to be a sensitive, specific and rapid method for detecting toxigenic strains of C. difficile in feces . Use of the method in diagnostic laboratories may require the develop ment of improved detection and verification systems for the amplified gene fragment.