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PubMed1993

Detection of toxigenic Clostridium difficile in stool specimens by the polymerase chain reaction.

Polymerase chain reaction (PCR) amplification of a segment of the toxin A gene was used to detect toxigenic Clostridium difficile directly from stool specimens of patients with antibiotic-associated diarrhea. Although PCR-inhibitory substances were recognized in DNA prepared from stool specimens, the inhibitory substances were eliminated by using an ion-exchange column after phenol-chloroform extraction. Eventually, 39 stool specimens were evaluated by PCR. PCR results for detection of toxigenic C. difficile were in complete agreement with cell culture assay results; all 12 PCR-positive stool specimens were positive by cytotoxin assay, and all 27 PCR-negative specimens were negative by cytotoxin assay. Toxigenic C. difficile was cultured from all PCR-positive specimens. These results suggest that PCR amplification may be an effective method for laboratory diagnosis of C. difficile-associated diarrhea and colitis.

Bacterial Toxins↗

PubMed1994

Wide diversity of Clostridium difficile types at a tertiary referral hospital.

Nosocomial Clostridium difficile infection was investigated at a hospital with 15 cases of C. difficile diarrhea per 1000 discharges. From January 1991 to May 1991, patients admitted or transferred to five wards or units had weekly rectal swabs taken for culture; in addition, all cytotoxin-positive stools were cultured. Restriction enzyme analysis (REA) was used for molecular typing. Among 205 isolates from 39 patients with C. difficile diarrhea and 67 asymptomatically colonized, 55 distinct REA banding patterns were identified. Evidence for patient-to-patient transmission was limited, in that numerous strains were found even among clustered cases of diarrhea. Patients who acquired C. difficile in the community or other hospitals constituted 32% of culture-positive patients and contributed 44% of the REA types. Diversity of C. difficile strains was in part the result of patients acquiring C. difficile in the community or other hospitals. High incidences of nosocomial C. difficile diarrhea do not necessarily indicate clonal epidemics.

Aged↗

PubMed1998

Clinical impact and associated costs of Clostridium difficile-associated disease.

Toxin-producing Clostridium difficile is the commonest cause of nosocomial diarrhoea and, as such, poses a major problem in our hospitals. The main population susceptible to disease is the elderly, for reasons that remain unclear. By contrast, carriage rates in neonates are high, but disease is low. The organism also has a major clinical impact in the immunosuppressed host, patients undergoing surgery (especially gastrointestinal) and those with severe underlying disease and longer hospital stay. Other interventions with high-risk associations are enemas, nasogastric and gastrostomy tubes and anti-peristaltic drugs. Data on the associated costs of C. difficile diarrhoea are not freely available, but one estimate is that in an average-sized district general hospital, 100 cases of C. difficile infection can be expected each year with an extra annual cost of 400,000 pound sterling and 2100 lost bed days.

Age Factors↗

PubMed2007

Update on Clostridium difficile associated disease.

PURPOSE OF REVIEW: The aim of this article is to report recent changes in the epidemiology of Clostridium difficile associated disease. RECENT FINDINGS: An epidemic of Clostridium difficile associated disease in Quebec was associated with a threefold increase in incidence and a sharp increase in fatalities. Strain typing of C. difficile isolates from the involved hospitals revealed that 82% were of a single strain (NAP1/027). This strain was found to produce greater than 10 times as much toxin A and toxin B as historic isolates and has been identified in many institutions throughout North America and Europe. Frequent nosocomial use of fluoroquinolones may encourage the spread of this strain as it is fluoroquinolone resistant. An increased rate of community-acquired Clostridium difficile-associated disease has also been noted and, in some cases, without prior antibiotic exposure. Although some studies have suggested an increased failure rate of metronidazole in Clostridium difficile associated disease, it remains the recommended first line treatment for uncomplicated cases. Other antibiotics, a toxin binder, probiotic agents and a vaccine are being tested in clinical trials for efficacy in prevention and treatment of Clostridium difficile associated disease. SUMMARY: The recent increase in the incidence and severity of Clostridium difficile associated disease may be related, at least in part, to the emergence of a highly virulent, fluoroquinolone-resistant, NAP1/027 strain.

Clostridioides difficile↗

PubMed1996

Cell surface properties of Clostridium difficile: haemagglutination, relative hydrophobicity and charge.

Five well characterised strains of Clostridium difficile of differing virulence and two Escherichia coli strains, a verotoxigenic O157:H7 isolate and a urinary isolate, were examined for cell surface hydrophobicity and charge, and haemagglutinating ability. Phase partition in hexadecane or octan-1-ol was similar for C. difficile and E. coli, as was retention by hydrophobic interaction chromatography (HIC), indicating moderate hydrophobicity. The salt agglutination test showed E. coli to be hydrophobic and C. difficile to be hydrophilic. Relative hydrophobicity determined by HIC when charge effects were not nullified, i.e., to reflect more closely conditions in vivo, showed C. difficile to bind less well. Growth of C. difficile in caecal emulsions to simulate conditions in vivo did not alter the cell surface hydrophobicity. The phase partition method for charge determination indicated that E. coli and C. difficile had a net negative charge, although this was weaker for C difficile than E. coli. However, although E. coli exhibited a net negative charge as determined by immuno-gold electronmicroscopy (IGEM), in keeping with the results of the phase partition method, C. difficile was shown to be predominantly positively charged by IGEM, and by movement in a charged field as determined by paper electrophoresis and a novel method based on light microscope observation. A cell-wall deficient mutant of C. difficile was weakly positively charged, showing that most of the charge resides in the cell wall.

Cell Membrane↗

PubMed2005

Bovine antibody-enriched whey to aid in the prevention of a relapse of Clostridium difficile-associated diarrhoea: preclinical and preliminary clinical data.

In a pilot study, the feasibility of immune whey protein concentrate (40%; immune WPC-40) to aid the prevention of relapse of Clostridium difficile diarrhoea was evaluated. Immune WPC-40 was made from milk after immunization of Holstein-Frisian cows with C. difficile-inactivated toxins and killed whole-cell C. difficile. Immune WPC-40 contained a high concentration of specific sIgA antibodies, and was effective in neutralizing the cytotoxic effect of C. difficile toxins in cell assays in vitro. Immune WPC-40 conferred protection from otherwise lethal C. difficile-associated caecitis in hamsters. To obtain preliminary data in humans, 16 patients (10 male; median 57 years) with toxin- and culture-confirmed C. difficile diarrhoea were enrolled in an uncontrolled cohort study. Nine had a history of relapsing C. difficile diarrhoea. After completion of standard antibiotic treatment, the patients received immune WPC-40 TID for 2 weeks; it was well tolerated and no treatment-related adverse effects were observed. In all but one case, C. difficile toxins had disappeared from the faeces upon completion of treatment. During a follow-up period of median 333 days (range 35 days to 1 year), none of the patients had suffered another episode of C. difficile diarrhoea. These preliminary data suggest that immune whey protein concentrate-40 may be of help in the prevention of relapse of C. difficile diarrhoea.

Animals↗

PubMed2003

Identification and characterization of a fibronectin-binding protein from Clostridium difficile.

A 68 kDa fibronectin-binding protein (Fbp68) from Clostridium difficile displaying significant homology to several established or putative Fbps from other bacteria was identified. The one-copy gene is highly conserved in C. difficile isolates. Fbp68 was expressed in Escherichia coli in fusion with glutathione S-transferase; the fusion protein and the native Fbp68 were purified. Immunoblot analysis and cell fractionation experiments revealed that Fbp68 is present on the surface of the bacteria. Far-immuno dot-blotting demonstrated that Fbp68 was capable of fixing fibronectin. Indirect immunofluorescence and ELISA were employed to demonstrate that C. difficile could bind both soluble and immobilized fibronectin. With competitive adherence inhibition assays it was shown that antibodies raised against Fbp68 partially inhibited attachment of C. difficile to fibronectin and Vero cells. Furthermore, Vero cells could fix purified membrane-immobilized Fbp68. Thus Fbp68 appears to be one of the several adhesins identified to date in C. difficile.

Adhesins, Bacterial↗

PubMed1992

Separate isolation of Clostridium difficile spores and vegetative cells from the feces of newborn infants.

A modified taurocholate-cefoxitin-cycloserine-fructose agar medium, pH 5.5, on which vegetative cells alone could grow, was newly devised for separate isolation of Clostridium difficile vegetative cells and spores from feces. The ratio of C. difficile-positive feces from healthy newborn infants younger than 10 days of the age was 30.8%, and 93.3% of feces from healthy infants older than 20 days were positive for C. difficile. C. difficile spores alone were detected in twenty-one samples (75%) of C. difficile-positive Twenty-eight specimens. Only 10.7% (3/28) C. difficile vegetative cells alone were detected. C. difficile spores alone were detected in one of nine healthy adults. These collective results offer potential explanations for high frequent isolations of C. difficile from newborn infants without occurrence of pseudomembranous colitis.

Bottle Feeding↗

PubMed1997

Clostridium difficile associated with acute colitis in mature horses treated with antibiotics.

Clostridium (C.) difficile, or its cytoxin, was demonstrated in faecal samples from 10 of 25 (40%) mature horses investigated with acute colitis treated primarily with antibiotics for disorders other than diarrhoea. C. difficile was not found in faecal samples from 140 horses without signs of enteric disorders, from 21 nondiarrhoeic horses treated with antibiotics, nor from 22 horses with colitis untreated with antibiotics. Except for C. difficile neither Salmonella nor any other investigated intestinal pathogen was isolated in any of the diarrhoeic horses. The findings strongly support some earlier reports that C. difficile is associated with acute colitis in mature horses treated with antibiotics. Of the 10 horses, 4 proved positive for C. difficile both in culture and in the cytotoxin test, 4 in culture only and 2 only in the cytotoxin test. Eight of 10 horses positive for C. difficile were or had recently been hospitalised, indicating that C. difficile may be a nosocomial infection in horses. All horses positive for C. difficile were treated with beta-lactam antibiotics.

Acute Disease↗

PubMed2005

Suspected Clostridium difficile-associated hemorrhagic diarrhea in a 1-week-old elk calf.

Clostridium difficile-associated diarrhea was suspected in a 1-week-old elk (Cervus elaphus) calf. The isolation of a toxigenic strain of C. difficile from a diarrheic fecal sample, along with exclusion of other enteropathogens, formed the basis of this presumptive diagnosis. Further study is indicated to evaluate the role of C. difficile in neonatal diarrhea in elk.

Animals↗

PubMed2006

High unmet-need infectious disease opportunities.

Despite advances in pharmaceutical science, considerable unmet medical needs exist in the field of anti-infective therapies. While pharmaceutical companies have focused research efforts on a few selected areas of the field, new microbial diseases have steadily increased in prevalence. Three emerging areas in the field of anti-infectives now represent large commercial opportunities: Gram-negative infections, community-acquired methicillin-resistant Staphylococcus aureus, and Clostridium difficile-associated diarrhea. Dedication and extensive investment by pharmaceutical companies is required to battle against these emerging and deadly microbial infections.

Anti-Infective Agents↗

PubMed1995

[Clostridium difficile infection. The nosocomial aspect].

Clostridium difficile infection is discussed relatively rarely as a nosocomial problem in Norway. Epidemiological studies have shown Clostridium difficile diarrhoea to be frequently an antibiotica-associated nosocomial disease. We describe a material consisting of 22 patients with Clostridium difficile infection from a county hospital in Norway. The inclusion criteria were a clinical disease with diarrhoea and a positive test for Clostridium difficile toxin A and/or B in the faeces. 11 patients were or had been hospitalized for a short period in one specific medical ward when diagnosed. These and five other patients probably had a hospital-acquired Clostridium difficile infection. We discuss the nosocomial aspect and the clinical characteristics of the disease in our material.

Adult↗

PubMed1993

[Antibiotic-induced diarrhea and pseudomembranous colitis].

Although not all antibiotic-induced diarrhoea is pseudomembranous, and not all pseudomembranous colitis is antibiotic-induced, by far the greater proportion of intestinal lesions and diarrhoea occurring in conjunction with Clostridium difficile infection are attributed to previous antibiotic treatment. The article consists in a review of pathogenic, diagnostic, treatment and nosocomial aspects.

Anti-Bacterial Agents↗

PubMed2002