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PubMed2002

High prevalence of toxin A-negative toxin B-positive Clostridium difficile in hospitalized patients with gastrointestinal disease.

The incidence of Clostridium difficile toxin A-negative toxin B-positive in hospitalized patients with severe gastrointestinal disease was evaluated. Of 530 stool specimens tested in parallel by two immunoassay tests, Tox A and Tox A/B (TechLab, Inc., Blacksburg, VA), 422 produced negative results on both tests. One hundred eight specimens (20.4%) tested positive by Tox A/B assay, and only 47 of them were also positive by Tox A. The 61 specimens with discrepant results were confirmed to be positive for toxin B by tissue culture cytotoxicity assay. Furthermore, 3 of the 422 specimens that were negative by enzyme immunoassay tested positive by cytotoxicity assay. The sensitivity and specificity of the Tox A/B test were 95.3% and 100%, respectively, with negative and positive predictive values of 99.3% and 100%, respectively, and a correlation rate of 99.4%. The high prevalence (56.5%) of specimens from symptomatic patients having detectable toxin B, but undetectable toxin A emphasizes the importance of using diagnostic tests that include toxin B. Furthermore, our data support the potential role of toxin B in the pathogenesis of toxigenic Clostridium difficile.

Bacterial Proteins↗
PubMed2003

Evaluation of the Clearview Clostridium difficile Toxin A Test and various selective culture media in comparison with the cytotoxin assay for the diagnosis of Clostridium difficile-associated diarrhoea.

AIM: Clostridium difficile is the major pathogen associated with nosocomial diarrhoea. We evaluated the performances of a commercially available toxin A enzyme immunoassay (EIA; Clearview C. difficile Toxin A Test), culture and tissue culture cytotoxin assay in the diagnosis of C. difficile-associated diarrhoea. METHODS: Comparative test performance was determined from data obtained from 166 faecal samples. The initial analysis compared the performance of toxin A EIA and culture with that of cytotoxin assay, this being defined as a 'laboratory gold standard'. A second analysis compared the individual performance of the toxin A EIA, culture and cytotoxin assay using a combined clinical and laboratory diagnostic assessment as a 'clinical gold standard'. In a parallel, study three selective culture media were compared. RESULTS: From the initial analysis, the sensitivity and specificity of the methods were, respectively, 84.6 and 65.4% for the toxin A EIA, and 38.5 and 93.5% for culture. From the second analysis, the sensitivity and specificity of the methods were, respectively, 100 and 67.5% for the toxin A EIA, 63.6 and 96.7% for culture and 72.7 and 98.0% for cytotoxin assay. Media containing d-cycloserine 250mg/L and cefoxitin 8mg/L performed best, growing 88.2% of the isolates. CONCLUSION: The toxin A EIA we evaluated had poor specificity in the diagnosis of C. difficile-associated diarrhoea. We conclude that in our laboratory the combination of culture and cytotoxin assay is a preferred approach to the diagnosis of C. difficile-associated diarrhoea.

Bacterial Toxins↗
PubMed1994

Evaluation of an enzyme immunoassay kit for the detection of Clostridium difficile enterotoxin.

The Premier Clostridium difficile toxin A enzyme immunoassay (EIA) kit was evaluated for the detection of C. difficile enterotoxin in fecal samples. A total of 314 samples was tested by culture, cytotoxin detection and EIA kit. Compared to a combined culture/cytotoxin result the Premier EIA kit had a sensitivity of 88.3%, a specificity of 100%, a predictive value positive of 100% and a predictive value negative of 87.4%. Test results were available within 3 hrs providing a rapid and reliable means of detecting C. difficile enterotoxin.

Clostridioides difficile↗
PubMed2001

Clostridium difficile small bowel enteritis occurring after total colectomy.

Clostridium difficile infection is usually associated with antibiotic therapy and is almost always limited to the colonic mucosa. Small bowel enteritis is rare: only 9 cases have been previously cited in the literature. This report describes a case of C. difficile small bowel enteritis that occurred in a patient after total colectomy and reviews the 9 previously reported cases of C. difficile enteritis.

Adult↗
PubMed2003

Activity of three disinfectants and acidified nitrite against Clostridium difficile spores.

OBJECTIVE: To identify environmentally safe, rapidly acting agents for killing spores of Clostridium difficile in the hospital environment. DESIGN: Three classic disinfectants (2% glutaraldehyde, 1.6% peracetyl ions, and 70% isopropanol) and acidified nitrite were compared for activity against C. difficile spores. Four strains of C. difficile belonging to different serogroups were tested using a dilution-neutralization method according to preliminary European Standard prEN 14347. For peracetyl ions and acidified nitrite, the subjective cleaning effect and the sporicidal activity was also tested in the presence of organic load. RESULTS: Peracetyl ions were highly sporicidal and yielded a minimum 4 log10 reduction of germinating spores already at short exposure times, independent of organic load conditions. Isopropanol 70% showed low or no inactivation at all exposure times, whereas glutaraldehyde and acidified nitrite each resulted in an increasing inactivation factor (IF) over time, from an IF greater than 1.4 at 5 minutes of exposure time to greater than 4.1 at 30 minutes. Soiling conditions did not influence the effect of acidified nitrite. There was no difference in the IF among the 4 strains tested for any of the investigated agents. Acidified nitrite demonstrated a good subjective cleaning effect and peracetyl ions demonstrated a satisfactory effect. CONCLUSIONS: Cidal activity was shown against C. difficile spores by glutaraldehyde, peracetyl ions, and acidified nitrite. As acidified nitrite and peracetyl ions are considered to be environmentally safe chemicals, these agents seem well suited for the disinfection of C. difficile spores in the hospital environment.

Clostridioides difficile↗
PubMed1994

Genotyping of Clostridium difficile isolates.

Arbitrarily primed polymerase chain reaction (AP-PCR) was used to genotype Clostridium difficile isolates from various sources. Four major molecular types were identified among strains from the American Type Culture Collection previously typed by serogroup and from isolates from patients at the University of California, Davis Medical Center, from a patient at a Utah institution, and from the environment. These groups contained subgroups that displayed, in addition to the common group bands, at least one unique band. Two strains isolated from patients at our institution had the same DNA banding patterns. These patients were hospitalized during the same period, raising the possibility of cross-infection through hospital contact or another common source. These results suggest that this AP-PCR approach will be useful in epidemiologic studies of C. difficile infections.

Base Sequence↗
PubMed1994

Characterization of the genetic basis of antibiotic resistance in Clostridium difficile.

By using dot blot hybridization, 69 of 102 Clostridium difficile isolates (68%) from the United States and other countries hybridized with at least one of nine DNA probes for erythromycin (Erm), tetracycline (Tet) or chloramphenicol (Cat) resistance determinants. The distribution of individual determinants in descending order of frequency was: Tet M, 32%; Erm Q, 25%; Erm FS, 18%; Tet P, 15%; Tet K, 15%; Cat P, 15%; Cat Q, 12%; Erm BP, 11%; Tet L, 7%. This is the first report of Tet P being carried by C. difficile and hitherto Erm FS has only been found within the genus Bacteriodes, while neither Tet K nor Tet L have been previously identified among the genus Clostridia. Eighteen percent of the hybridizing isolates carried multiple determinants coding for the same phenotype. A higher frequency of resistance genes was associated with prior exposure to antimicrobial agents, cytotoxin production and diarrhoea. Isolates recovered from bone marrow transplant patients carried significantly fewer antibiotic resistance genes than did those from immunocompetent general medicine patients. However, this may be due to the fact that each was located at a different site. Antibiotic resistance determinants may play a role in the virulence associated with C. difficile.

Clostridioides difficile↗