AIMS: A set of five tests were developed and tested for their ability to confirm the identity of C difficile colonies within 30 minutes. METHODS: The relevant substrates were incorporated into four filter paper squares attached to a plastic carrier (Diffstrip), five enzymes/products (prolyl aminopeptidase, galactosidase, leucine aminopeptidase, acid phosphatase and indole). The strips were inoculated, incubated for 20 minutes, and reagents added. RESULTS: 96.4% (212 of 220) strains of C difficile were immediately differentiated from 51 other Clostridium spp tested. The remaining 3.6% (eight of 220) of C difficile isolates produced a reaction pattern similar to some of the Clostridium sporogenes tested and required additional tests. None of the other Clostridium spp tested produced reaction patterns similar to C difficile. CONCLUSION: The Diffstrip allowed colonies of C difficile to be confirmed within 30 minutes for 96.4% of isolates, with less than 4% requiring any additional tests. No strains of C difficile were misidentified and no strains of other Clostridium spp tested were misidentified as C difficile.
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