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PubMed2006

Prevalence of Clostridium difficile in the environment in a rural community in Zimbabwe.

Clostridium difficile has been shown to be a nosocomial pathogen associated with diarrhoea and pseudomembranous colitis in hospitalised patients, but very little is known about its prevalence outside the hospital environment. The aim of this study was to determine the prevalence of C. difficile in faeces of domestic animals, soil and drinking water in a rural community. Water, animal faeces and soil were collected from homesteads in a rural community and the samples were cultured for C. difficile. Clostridium difficile isolates that produced toxins A or B were tested for their susceptibility to antimicrobial drugs. Clostridium difficile was isolated from 37.0% of 146 soil samples, 17.4% of 115 chicken faeces samples, 6.0% of 234 water samples and 4.3% of 161 faecal samples of other animals. Some of the C. difficile isolates from chickens (55.0%), soil (66.7%) and water (14.3%) were toxigenic. All toxigenic isolates were susceptible to metronidazole, vancomycin, doxycycline, chloramphenicol and tetracycline and all were resistant to cefotaxime, gentamicin, ciprofloxacin, norfloxacin and nalidixic acid. The results of the present study suggest that chickens kept by villagers are an important reservoir of C. difficile, which may act as a source of human infection.

Animals↗
PubMed1996

A receptor decoy inhibits the enterotoxic effects of Clostridium difficile toxin A in rat ileum.

BACKGROUND & AIMS: Clostridium difficile toxin A causes secretion and intestinal inflammation in rodents by binding to a specific trisaccharide Gal alpha 1-3Gal beta 1-4 GlcNAc on enterocyte receptors. The purpose of this study was to explore the ability of Synsorb 90 (Synsorb Biotech Inc., Calgary, Alberta, Canada), and inert support carrying this trisaccharide, to bind toxin A in vitro and to inhibit its enterotoxic effects in vivo. METHODS: Binding of [3H]toxin A to Synsorb 90, Synsorb 83 (beta-mannose attached), and Chromosorb P (inert support with no sugar attached) (Synsorb Biotech Inc.) was measured. The inhibitory effects of these compounds on toxin A-mediated fluid secretion, mannitol permeability, and histological damage were measured in ileal loops in vivo. RESULTS: Toxin A showed specific binding to Synsorb 90, bearing the specific trisaccharide that binds toxin A, but not to Synsorb 83 or to Chromosorb P. Pretreatment of rats with Synsorb 90 by gavage (200 mg/kg body wt), but no Synsorb 83 or Chromosorb P at the same doses, dramatically reduced toxin A-associated fluid secretion and permeability. CONCLUSIONS: An immobilized toxin A receptor sequesters toxin A in the intestinal lumen and inhibits its effects of ileal mucosa. These results suggest a potential use for this agent in treating patients with C. difficile colitis.

Animals↗
PubMed2000

The rise of Clostridium difficile: the effect of length of stay, patient age and antibiotic use.

Hospitals in the UK have recently seen a marked increase in C. difficile for reasons which are unclear. Reduced standards of hygiene, increasingly elderly patients, greater cephalosporin use and longer hospital stay have been suggested. We retrospectively studied all cases of C. difficile diarrhoea at Princess Margaret Hospital, Swindon, over two years. Cephalosporins, patient age and LOS appeared unrelated to the rise in C. difficile; penicillins and macrolides were related. Our policy of using amoxycillin and clarithromycin for community-acquired pneumonia coincided with this study and may explain the observed rise in C. difficile.

Age Factors↗
PubMed2000

Asymptomatic carriage of Clostridium difficile and serum levels of IgG antibody against toxin A.

BACKGROUND: Clostridium difficile infection can result in asymptomatic carriage, mild diarrhea, or fulminant pseudomembranous colitis. We studied whether antibody responses to C. difficile toxins affect the risks of colonization, diarrhea, and asymptomatic carriage. METHODS: We prospectively studied C. difficile infections in hospitalized patients who were receiving antibiotics. Serial stool samples were tested for C. difficile colonization by cytotoxin assay and culture. Serum antibody (IgA, IgG, and IgM) levels and fecal antibody (IgA and IgG) levels against C. difficile toxin A, toxin B, and nontoxin antigens were measured by an enzyme-linked immunosorbent assay (ELISA). RESULTS: Of 271 patients, 37 (14 percent) were colonized with C. difficile at the time of admission, 18 of whom were asymptomatic carriers. An additional 47 patients (17 percent) became infected in the hospital, 19 of whom remained asymptomatic. The baseline antibody levels were similar in the patients who later became colonized and those who did not. After colonization, those who became asymptomatic carriers had significantly greater increases in serum levels of IgG antibody against toxin A than did the patients in whom C. difficile diarrhea developed (P<0.001). The adjusted odds ratio for diarrhea was 48.0 (95 percent confidence interval, 3.4 to 678) among patients with colonization who had a serum level of IgG antibody against toxin A of 3.00 ELISA units or less, as compared with patients with colonization who had a level of more than 3.00 ELISA units. CONCLUSIONS: We find no evidence of immune protection against colonization by C. difficile. However, after colonization there is an association between a systemic anamnestic response to toxin A, as evidenced by increased serum levels of IgG antibody against toxin A, and asymptomatic carriage of C. difficile.

Aged↗
PubMed1999

An antibiotic policy associated with reduced risk of Clostridium difficile-associated diarrhoea.

BACKGROUND: antibiotic-associated diarrhoea caused by Clostridium difficile is increasing in hospitals, and older people are at particular risk. OBJECTIVE: to establish whether reducing patient exposure to injectable third-generation cephalosporins by substituting alternative antibiotics can produce a cost-effective reduction in the incidence of antibiotic-associated diarrhoea. DESIGN: we prospectively investigated 2157 patients admitted to the department of elderly medicine in the year before introduction of antibiotic restrictions and 2037 patients admitted in the following year. Patients admitted to other wards, where antibiotic prescribing was unchanged, acted as controls. SETTING: a 900-bed teaching hospital in Cambridge, UK. MEASUREMENTS: use and cost of injectable antibiotics prescribed in the department of elderly medicine and the other wards studied; occurrence of C. difficile-associated diarrhoea. RESULTS: in the wards for older people, consumption of injectable cephalosporins fell by 92% (compared with 8% on other wards) and cases of C. difficile-associated diarrhoea fell from 98 to 45 (cases in other wards rose from 213 to 253; P < 0.001). The Pound Sterling 8062 increase in injectable antibiotic costs on the elderly wards were offset by the release of 1087 wasted bed-days attributable to the 53 fewer cases, with potential savings of Pound Sterling 212,000. CONCLUSIONS: restricting the consumption of injectable third-generation cephalosporins is a cost-effective method of reducing the incidence of C. difficile-associated diarrhoea.

Aged↗
PubMed1999

Detection of Clostridium difficile toxin A by reversed passive latex agglutination.

A reversed passive latex agglutination (RPLA) assay for detecting Clostridium difficile toxin A is presented. Purified monoclonal antibody (mAb 37B5) was used for latex sensitization. The culture supernatants of 93 strains of C. difficile were tested by RPLA assay and the results compared with those of a commercially available latex agglutination test, PCR and cytotoxin assay with Vero cells. There was agreement between RPLA, cytotoxicity and PCR assays, but 29 strains were positive in the RPLA assay while 35 were positive in the cytotoxicity test and PCR using primer pair NK3-NK2 directed to the nonrepeating portion of the C. difficile toxin A gene. The 6 cytotoxic but RPLA-negative strains were demonstrated to be toxin A-negative/toxin B-positive strains in the PCR assay by using primer pair NK11-NK9 directed to the repeating portion of the C. difficile toxin A gene. There were no cross-reactions with culture supernatants of the other clostridial strains except for two strains of C. sordelli that produced hemorrhagic toxin (which is immunologically related to C. difficile toxin A).

Agglutination Tests↗
PubMed2005

Clostridium difficile associated severe enterocolitis: a feature of Hirschsprung's disease in a neonate presenting late.

We report a case of Hirschsprung's disease presenting late in the newborn period with severe enterocolitis and Clostridium difficile toxin detectable in the stool. Enterocolitis associated with Hirschsprung's disease is a potentially fatal complication that may occur at presentation or later in the life of an affected child. Its association with toxin-producing Clostridium difficile growth in the bowel may be under recognized, especially in newborns.

Bacterial Toxins↗
PubMed1993

Use of an enzyme-linked immunoassay for Clostridium difficile serogrouping.

An enzyme-linked immunoassay (ELISA) with 11 Clostridium difficile serogroup-specific antisera was applied for serogrouping of C. difficile colonies from 314 consecutive positive fecal samples. Two hundred forty-nine strains (79%) were correctly serogrouped, 57 (18%) belonged to serogroups other than the 11 which were evaluated and gave a negative reaction with all antisera, and 8 isolates (2.5%) did not react with their corresponding antisera. ELISA is a rapid and reliable method for serogrouping C. difficile and should allow for the automation of this procedure.

Animals↗
PubMed2001

Evaluation of biosite triage Clostridium difficile panel for rapid detection of Clostridium difficile in stool samples.

One hundred two stool samples were tested by both the rapid Triage Clostridium difficile Panel (Triage Panel) and the cytotoxin cell culture assay. Five samples positive by both the C. difficile toxin A (Tox A) and common antigen components of the Triage Panel had cytotoxin titers of > or =10,000. Twenty-three samples were Triage Panel Tox A negative but common antigen positive. Ten of these had cytotoxin titers of 10 to 1,000, but 13 were cytotoxin negative. Bacterial isolates obtained from 8 of these 13 specimens were analyzed for Tox A and B genes by PCR, and only two contained toxigenic bacteria. Thus, the majority of samples positive only for C. difficile common antigen contained nontoxigenic bacteria. A Triage Panel Tox A-positive result indicated a sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of 33.3, 100, 100, and 88.2%, respectively. A Triage Panel common antigen-positive result indicated a sensitivity, specificity, PPV, and NPV of 100, 82.7, 53.6, and 100%, respectively. The high NPV of the Triage Panel common antigen, together with rapid reporting of results, should prove useful in avoiding unnecessary use of contact precautions and antibiotic treatment for C. difficile-negative patients. However, with Triage Panel common antigen-positive patients, a sensitive cytotoxin assay should be used to distinguish true cytotoxin-positive patients from C. difficile carriers.

Antigens, Bacterial↗
PubMed2004

Performance of the TechLab C. DIFF CHEK-60 enzyme immunoassay (EIA) in combination with the C. difficile Tox A/B II EIA kit, the Triage C. difficile panel immunoassay, and a cytotoxin assay for diagnosis of Clostridium difficile-associated diarrhea.

We compared a recently marketed enzyme immunoassay for glutamate dehydrogenase (GDH), TechLab's C. DIFF CHEK-60 (TL-GDH), in combination with the C. difficile Tox A/B II enzyme immunoassay (Tox-A/B) with (i) the Triage C. difficile test, which detects both GDH (TR-GDH) and toxin A (TR-Tox-A); (ii) an in-house cytotoxin assay (C-Tox); and (iii) stool cultures for C. difficile. All C. difficile isolates were tested for the presence of the toxin genes by PCR. If a toxin gene-positive strain of Clostridium difficile was recovered and a toxin was detected by any method, the result was considered to be truly positive. Eighty-seven of 93 and 79 of 93 C. difficile culture-positive samples were also TL-GDH and TR-GDH positive, respectively. No test was able to detect toxin in all samples with true-positive results. Tox-A/B and TR-Tox-A in combination with the GDH detection tests and C-Tox were able to identify 52 and 50 samples with true-positive results. Tox-A/B and TR-Tox-A would have missed 15 and 31% of cases of C. difficile-associated diarrhea, respectively, if used alone.

Bacterial Proteins↗