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PubMed1992

New rapid identification test for Clostridium difficile.

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.

Bacterial Typing Techniques↗
PubMed1994

Evaluation of three commercial enzyme immunoassay kits for detecting faecal Clostridium difficile toxins.

The detection of faecal cytotoxicity using tissue culture was compared with three commercial Clostridium difficile enzyme immunoassay (EIA) kits; Premier C difficile toxin A (Meridian Diagnostic, Inc.); CD-TOX C difficile toxin A (Porton Cambridge); and Cytoclone A+B EIA (Cambridge Biotech Corporation). Of 160 faecal samples examined by all four methods, 52 (32.5%) were cytotoxic, 44 (27.5%) were positive by Premier, 48 (30%) by CD-TOX EIA, and 50 (31.3%) with Cytoclone. When compared with detection of cytotoxicity by tissue culture assay, the following performance indices were obtained: Premier, sensitivity 84.1%, specificity 99.1%, positive predictive value (PPV) 97.8%, negative predictive value (NPV) 93%; CD-TOX, sensitivity 92.3%, specificity 88.0%, PPV 78.7%, NPV 95.9%; Cytoclone, sensitivity 96.2%, specificity 93.5%, PPV 87.7%, NPV 98.1%. EIA results were available within three hours, whereas the results of the cytotoxin assay were available after 24-48 hours. All three kits provided satisfactory results and, although relatively expensive, all could be used in the laboratory effectively to screen for diarrhoeal disease associated with C difficile.

Clostridioides difficile↗
PubMed2002

Recurrent Clostridium difficile colitis. Tackling a tenacious nosocomial infection.

C difficile infection recurs in about 20% of previously treated hospitalized patients. The elderly and patients with underlying colonic disease who have recently used antibiotics are at high risk. Signs and symptoms include diarrhea, abdominal pain, and leukocytosis. Diagnosis is dependent on a high degree of clinical suspicion and ELISA testing of a stool sample for toxins. Recurrence is thought to be due to the persistence of C difficile spores. Treatment can be difficult. Oral vancomycin or metronidazole for 10 to 14 days may be helpful as first-line therapy. Tapering the dose over 1 month helps destroy the spores while enabling the normal colonic flora to regrow. Probiotics, such as lactobacillus GG and S boulardii, are being developed to help restore normal colonic flora. Immunization may help prevent C difficile infection in the first place.

Clostridioides difficile↗
PubMed2005

[Comparison of rapid tests of toxin A and glutamate dehydrogenase and culture for detection of Clostridium diffcile].

Clostridium difficile is a major cause of antibiotics-associated diarrhea (AAD), and accounts for 15-20% of all the cases. Especially, AAD caused by C. difficile is called as C. difficile-associated diarrhea (CDAD). Diagnosis of CDAD is made by identification of C. difficile in the feces obtained from the patients with diarrhea after administration of antibiotics. We herein compared 3 methods, detection of toxin A and glutamate dehydrogenase (GDH) as well as culture for C. difficile. Two hundred and thirty-two samples obtained from 148 patients with CDAD were investigated. Fifty-five (23.7%), 56 (24.1%), and 93 (40.1%) of the 232 samples were positive for toxin A, GDH, and culture, respectively. Sensitivity, specificity, predictive value of positive result, and predictive value of negative result of method of detection of toxin A were estimated to 53%, 96%, 90%, 75% and those of GDH were 56%, 99%, 98%, 78%. These results indicated that we should carefully interpret the results obtained by these methods, especially in the cases that showed negative for C. difficile.

Aged↗
PubMed1991

Clostridium difficile infection in adult hamsters.

Diarrhea was encountered in a group of adult female golden Syrian hamsters (Mesocricetus auratus) used for titrating the scrapie agent. Ninety percent of the cases occurred in animals over 210 days old even though animals of all age groups lived in the colony concurrently. The cause of diarrhea was investigated in both uninoculated animals and those receiving greater than a limiting dilution of scrapie infectivity, i.e., animals that were not expected to contract the experimental scrapie disease. Three forms of diarrhea were observed. The most commonly encountered was profuse and watery. A chronic form presented with semiformed, thin fecal material smearing the retroperitoneal region. Hemorrhagic diarrhea was observed rarely. Mortality was high among animals with acute watery or hemorrhagic diarrhea. Animals with semiformed soft stools were dehydrated, had a roughened hair-coat, and hunched back. Cardinal lesions were necrosis, inflammation, and mucosal hyperplasia of the cecum and colon and cholangiohepatitis with amyloid deposition. Diffuse renal amyloidosis was present in chronic cases. Toxigenic, cytotoxin B-positive Clostridium difficile was isolated from a majority of affected animals. Cytotoxin B was also present in cecal homogenates of diarrheic animals with C. difficile. The pathological and microbiologic findings indicated a typhlitis and colitis in adult hamsters that was associated with C. difficile infection.

Animals↗
PubMed1991

Capsule-like structures in Clostridium difficile strains.

Fourteen strains of Clostridium difficile, previously characterized by SDS-PAGE, were examined for the presence of surface structures. None of the strains were fimbriated but, when grown in the presence of glucose, all exhibited a thin ruthenium red-positive layer. Two strains, belonging to different electrophoretic groups, were also observed after treatment with homologous and heterologous antisera. The strain belonging to the electrophoretic group 2, usually associated with the disease, showed a 30-80nm thick capsule in ultrathin sections. The strains belonging to group 5, mainly observed in isolates from healthy children, exhibited a thinner polysaccharide layer (10-20 nm). No stabilization was observed when these strains were treated with heterologous antisera. This capsule-like material did not seem to be associated with adhesive properties of C. difficile strains.

Cell Wall↗
PubMed1991

[Generalized toxic symptoms caused by Clostridium difficile infection in a patient with Wilms' tumor].

This case report describes a 15 months-old patient with Wilms' tumor, who was treated with radical operation and chemotherapy. During hospitalization the patient suffered from recurrent lung infections, which were treated with appropriate antibiotics. Subsequently, the patient developed severe abdominal affection with daily melena, liver and kidney involvement, and generalized edema and hypertension. Cl. difficile and its cytotoxin was demonstrated in feces, and the patient was successfully treated with vancomycin. Although rare, Cl. difficile may cause severe generalised illness in children under two years of age, and it is important to examine any infant with abdominal symptoms for the occurrence of Cl. difficile, even though no diarrhoea is present.

Clostridioides difficile↗