New selective medium for isolating Clostridium difficile from faeces.

Aspinall, S T; Hutchinson, D. N.

doi:10.1136/jcp.45.9.812

Cited by 49 publications

(25 citation statements)

References 9 publications

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“…Our observation that plates with a reduced cycloserine concentration of 250 g/ml resulted in increased breakthrough growth of other organisms is consistent with those of previous studies (1,16). Aspinall and Hutchinson (1) found that norfloxacin at 12 g/ml in combination with moxalactam was as effective as cycloserine and cefoxitin for suppressing breakthrough growth of stool microflora.…”

Section: Discussionsupporting

confidence: 82%

“…Norfloxacin at 12 or 32 g/ml was evaluated as an alternative to D-cycloserine because it has been reported previously to be effective, when used in combination with moxalactam (latamoxef), for selective isolation of C. difficile (1). However, high levels of breakthrough growth of multiple organisms other than C. difficile from 30 toxin-positive stool samples were observed on CCFA plates in which norfloxacin had been substituted for D-cycloserine.…”

Section: Methodsmentioning

confidence: 99%

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Effective and Reduced-Cost Modified Selective Medium for Isolation of Clostridium difficile

Nerandzic

Donskey

2009

J Clin Microbiol

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Both for epidemiologic studies and for diagnostic testing, there is a need for effective, economical, and readily available selective media for the culture of Clostridium difficile. We have developed a reduced-cost substitute for cycloserine-cefoxitin-fructose agar (CCFA), which is an effective but expensive selective medium for C. difficile. The modified medium, called C. difficile brucella agar (CDBA), includes an enriched brucella base as a substitute for proteose peptone no. 2, and the concentration of sodium taurocholate has been reduced from 0.1% to 0.05%. To compare the sensitivities and selectivities of CDBA and CCFA, cultures for C. difficile were performed using stool samples from patients with C. difficile-associated disease. CDBA was as sensitive as CCFA for the recovery of C. difficile, with a similar frequency of breakthrough growth of stool microflora (25% versus 31%, respectively). A liquid formulation of the modified medium, termed C. difficile brucella broth (CDBB), stimulated rapid germination and outgrowth of C. difficile spores, at a rate comparable to that in cycloserine-cefoxitin-fructose broth. Our results suggest that CDBA and CDBB are sensitive, selective, and reduced-cost media for the recovery of C. difficile from stool samples.

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Section: Discussionsupporting

confidence: 82%

Section: Methodsmentioning

confidence: 99%

Effective and Reduced-Cost Modified Selective Medium for Isolation of Clostridium difficile

Nerandzic

Donskey

2009

J Clin Microbiol

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“…The culture method utilized in this study involved alcohol shock of stools, followed by culture on CDMN agar. CDMN agar has been demonstrated to yield improved recovery of C. difficile from stool specimens in comparison with cycloserinecefoxitin-fructose agar (CCFA) (19). Culture of all stool specimens was not performed in other studies evaluating the sensitivity and specificity of the Illumigene assay, and when culture was carried out, CCFA medium was generally used without any indication of prior alcohol shock treatment (10,11,17).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of an Algorithmic Approach in Comparison with the Illumigene Assay for Laboratory Diagnosis of Clostridium difficile Infection

et al. 2013

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The following three diagnostic algorithms were evaluated in comparison with the Illumigene assay as a stand-alone test for Clostridium difficile detection: glutamate dehydrogenase antigen screen (GDH) followed by toxin A/B antigen testing (Tox A/B) with the cell cytotoxicity assay for discordant specimens (algorithm 1), GDH followed by the Illumigene (algorithm 2), and GDH followed by Tox A/B with the Illumigene for discordant specimens (algorithm 3). A total of 428 stool specimens submitted to three clinical microbiology laboratories in Manitoba, Canada, for C. difficile detection between June 2011 and April 2012 were included in the study. The prevalence of C. difficile in the stool specimens was 14.7% (63/428) based on toxigenic culture (microbiologic reference standard). The sensitivity and specificity of the Illumigene for C. difficile detection were 73.0% and 99.7%, respectively. The corresponding sensitivities and specificities were 65.1% and 100.0% for algorithm 1, 68.3% and 100.0% for algorithm 2, and 69.8% and 100.0% for algorithm 3. Using algorithm 1, a cell cytotoxicity assay was required for toxin detection in 37% of positive tests, prolonging turnaround time. However, the predictive value of a positive test based on a clinical reference standard (all tests positive or cytotoxigenic culture positive and clinical disease on chart review) was slightly higher with algorithm 1 than with the Illumigene assay as a stand-alone test or as part of an algorithm (algorithms 2 and 3). Based on a reduction in turnaround time, simplicity, and acceptable sensitivity and specificity, we recommend algorithm 2 (screening with the GDH antigen test and confirmatory testing with the Illumigene). C lostridium difficile is an anaerobic, spore-forming Gram-positive bacillus that is capable of causing diarrhea mediated by the production of C. difficile toxins A and B (1-3). It is estimated that C. difficile infection (CDI) is responsible for 15 to 25% of cases of antibiotic-associated diarrhea (1). Severe CDI may result in the need for intensive care unit admission or even colectomy (4). Further, CDI is associated with an attributable mortality rate of approximately 5.7% (4). Related to both the frequency and severity of disease caused by C. difficile, it is imperative that diagnostic testing be sensitive, specific, and rapid, such that appropriate therapy may be instituted in a timely fashion (5).For diagnosis of CDI, many clinical microbiology laboratories currently use enzyme immunoassays (EIAs) that detect C. difficile toxins (1). These assays have the advantage of a rapid turnaround time. However, they demonstrate suboptimal sensitivity (5, 6). Algorithms relying on detection of the C. difficile glutamate dehydrogenase antigen (GDH) with follow-up toxin testing for positive specimens with a cell cytotoxicity assay demonstrate improved sensitivity (5, 7). However, the turnaround time may be increased by up to 48 h with this approach (5, 7). Additionally, recent data suggest that the cell cytotoxicity assay for toxin det...

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“…For measuring the bacterial count of C. difficile in faeces, stool pellets were suspended in PBS. The suspension was inoculated onto a selective medium containing cysteine hydrochloride, moxalactam and norfloxacin after 1 : 10 serial dilutions, and then cultivated anaerobically, as mentioned above (Aspinall & Hutchinson, 1992). For histological analysis, caecum was harvested on day 3 post-infection.…”

Section: Methodsmentioning

confidence: 99%

Endogenous IL-17 as a factor determining the severity of Clostridium difficile infection in mice

Nakagawa

Môri

Kajiwara

et al. 2016

Journal of Medical Microbiology

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Clostridium difficile infection (CDI) is a toxin-mediated intestinal disease. Toxin A, toxin B and binary toxin are believed to be responsible for the pathogenesis of CDI, which is characterized by massive infiltration of neutrophils at the infected intestinal mucosa. IL-17 is one of the cytokines that play critical roles in several inflammatory and immunological diseases through various actions, including promoting neutrophil recruitment. The aim of this study was to examine the role of this cytokine in CDI by employing IL-17 A and F double knockout (IL-17 KO) mice for the CDI model. We demonstrated that IL-17 KO mice were more resistant to CDI than WT mice using several factors, such as diarrhoea score, weight change and survival rate. Although the bacterial numbers of C. difficile in faeces were not different, the inflammatory mediator levels at the large intestine on day 3 post-infection were attenuated in IL-17 KO mice. Finally, we showed that infiltration of neutrophils, but not macrophages, in the large intestine was significantly decreased in IL-17 KO mice compared to WT mice. In conclusion, the data demonstrate that endogenous IL-17 may be a factor determining the severity of CDI in mice. Although the mechanism is totally unknown, IL-17-mediated inflammatory responses, such as cytokine/chemokine production and neutrophil accumulation, may be plausible targets for future investigations.

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New selective medium for isolating Clostridium difficile from faeces.

Cited by 49 publications

References 9 publications

Effective and Reduced-Cost Modified Selective Medium for Isolation of Clostridium difficile

Effective and Reduced-Cost Modified Selective Medium for Isolation of Clostridium difficile

Evaluation of an Algorithmic Approach in Comparison with the Illumigene Assay for Laboratory Diagnosis of Clostridium difficile Infection

Endogenous IL-17 as a factor determining the severity of Clostridium difficile infection in mice

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