C difficile agglutination test kit

To improve the sensitivity of C. difficile detection, we aliquoted each stool sample into two parts. One aliquot was pretreated with ethanol and then directly plated onto cycloserine cefoxitin fructose agar (Oxoid Ltd., Basingstoke, UK). The other was pre-cultured in cycloserine cefoxitin fructose medium supplemented with taurocholic acid and lysozyme for 48 h to enrich the vegetative cells prior to plating on cycloserine cefoxitin fructose agar. Culturing was performed at 35 °C for 48 h in anaerobic condition. Suspected C. difficile colonies were identified based on their odor and appearance, and confirmed using a latex agglutination test (C. difficile Agglutination Test Kit; Oxoid), gluD gene detection, and a toxin A & B test using a VIDAS Immunoanalyzer (Biomerieux, Marcy-l’Etoile, France).
To screen S. aureus isolates from stool samples, samples were plated on mannitol salt agar (Becton Dickinson) and cultured for 48 h at 35 °C in aerobic condition. Suspected S. aureus colonies were identified based on microbiological examination. MRSA was further confirmed by Kirby-Bauer disk diffusion assay (30 mg/liter cefoxitin) [11 ] and mecA gene detection using S. aureus ATCC25923 as a negative control.

Stool samples were collected from ICU patients within a set time period, plated onto C. difficile agar base supplemented with norfloxacin and moxalactam (Oxoid Ltd., Basingstoke, UK), and cultured anaerobically at 37 °C for 48–72 h. Colonies were identified by morphological features, latex agglutination test (C. difficile Agglutination Test Kit; Oxoid Ltd.), and gluD gene detection. Feces and C. difficile isolates were also subjected to toxin A/B detection by enzyme-linked fluorescence assay with a VIDAS automatic analyzer (Biomerieux, Marcy-l’Etoile, France) [22 (link)–24 (link)].