Comparison of culture based methods for the isolation of Clostridium difficile from stool samples in a research setting

Michelle Lister, Emma Stevenson, Daniela Heeg, Nigel P Minton, Sarah A Kuehne

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12 Citations (Scopus)
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Effective isolation of Clostridium difficile from stool samples is important in the research setting, especially where low numbers of spores/vegetative cells may be present within a sample. In this study, three protocols for stool culture were investigated to find a sensitive, cost effective and timely method of C. difficile isolation. For the initial enrichment step, the effectiveness of two different rich media, cycloserine-cefoxitin fructose broth (CCFB) and cycloserine-cefoxitin mannitol broth with taurocholate and lysozyme (CCMB-TAL) were compared. For the comparison of four different, selective solid media; Cycloserine-cefoxitin fructose agar (CCFA), Cycloserine-cefoxitin egg yolk agar (CCEY), ChromID C. difficile and tryptone soy agar (TSA) with 5% sheep's blood with and without preceding broth enrichment were used. As a means to enable differentiation between C. difficile and other fecal flora, the effectiveness of the inclusion of a pH indictor (1% Neutral Red), was also evaluated. The data derived indicated that CCFB is more sensitive than CCMB-TAL, however, the latter had an improved recovery rate. A broth enrichment step had a reduced sensitivity over direct plating. ChromID C. difficile showed the best recovery rate whereas CCEY egg yolk agar was the most sensitive of the four. The addition of 1% Neutral Red did not show sufficient colour change when added to CCEY egg yolk agar to be used as a differential medium. For a low cost, timely and sensitive method of isolating C. difficile from stool samples we recommend direct plating onto CCEY egg yolk agar after heat shock.

Original languageEnglish
Pages (from-to)226-229
Number of pages4
Early online date16 Jul 2014
Publication statusPublished - Aug 2014


  • Animals
  • Bacteriological Techniques
  • Clostridium difficile
  • Cost-Benefit Analysis
  • Culture Media
  • Feces
  • Mesocricetus
  • Sensitivity and Specificity
  • Time Factors
  • Comparative Study
  • Evaluation Studies
  • Journal Article
  • Research Support, Non-U.S. Gov't


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