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PubMed1995

Rapid polymerase chain reaction method for specific detection of toxigenic Clostridium difficile.

A rapid polymerase chain reaction (PCR) method for detection of toxigenic Clostridium difficile directly from fecal samples by amplification of toxin A gene fragments was investigated. The technique was applied to monitor the spread of the microorganism in a long-term care ward with a relatively high incidence of overt episodes of diarrhea. The PCR approach has several advantages over traditional methods, rapidly allowing the specific detection of toxigenic Clostridium difficile strains from stool samples in both symptomatic and asymptomatic subjects with toxigenic strains. This PCR method allows early detection of toxigenic Clostridium difficile and could thus represent a powerful tool for the surveillance of epidemics.

Clostridioides difficile↗
PubMed1990

Prospective, controlled study of vinyl glove use to interrupt Clostridium difficile nosocomial transmission.

PURPOSE: Despite recognition that Clostridium difficile diarrhea/colitis is a nosocomial infection, the manner in which this organism is transmitted is still not clear. Hands of health care workers have been shown to be contaminated with C. difficile and suggested as a vehicle of transmission. Therefore, we conducted a controlled trial of the use of disposable vinyl gloves by hospital personnel for all body substance contact (prior to the institution of universal body substance precautions) to study its effect on the incidence of C. difficile disease. PATIENTS AND METHODS: The incidence of nosocomial C. difficile diarrhea was monitored by active surveillance for six months before and after an intensive education program regarding glove use on two hospital wards. The interventions included initial and periodic in-services, posters, and placement of boxes of gloves at every patient's bedside. Two comparable wards where no special intervention was instituted served as controls. RESULTS: A decrease in the incidence of C. difficile diarrhea from 7.7 cases/1,000 patient discharges during the six months before intervention to 1.5/1,000 during the six months of intervention on the glove wards was observed (p = 0.015). No significant change in incidence was observed on the two control wards during the same period (5.7/1,000 versus 4.2/1,000). Point prevalence of asymptomatic C. difficile carriage was also reduced significantly on the glove wards but not on the control wards after the intervention period (glove wards, 10 of 37 to four of 43, p = 0.029; control wards, five of 30 to five of 49, p = 0.19). The cost of 61,500 gloves (4,505 gloves/100 patients) used was $2,768 on the glove wards, compared with $1,895 (42,100 gloves; 3,532 gloves/100 patients) on the control wards. CONCLUSIONS: Vinyl glove use was associated with a reduced incidence of C. difficile diarrhea and is indirect evidence for hand carriage as a means of nosocomial C. difficile spread.

Clostridioides difficile↗
PubMed2005

A modified pulsed-field gel electrophoresis (PFGE) protocol for subtyping previously non-PFGE typeable isolates of Clostridium difficile polymerase chain reaction ribotype 001.

A modified pulsed-field gel electrophoresis (PFGE) protocol was developed and applied to 50 isolates of the UK epidemic strain of Clostridium difficile, polymerase chain reaction (PCR) ribotype 001, to develop a PFGE-based subtyping scheme. This protocol overcame the inherent DNA degradation problems associated with typing this strain of C. difficile by this method, and whole genomic digestion with SmaI restriction enzyme yielded seven distinct and reproducible PFGE banding patterns. Modified PFGE is an appropriate method for subtyping C. difficile PCR ribotype 001 that could be used to improve epidemiological investigations.

Bacterial Typing Techniques↗
PubMed2006

Fatal enterocolitis in Asian elephants (Elephas maximus) caused by Clostridium difficile.

Two cases of fatal enteritis caused by Clostridium difficile in captive Asian elephants are reported from an outbreak affecting five females in the same zoo. Post mortem examination including histopathology demonstrated fibrinonecrotic enterocolitis. C. difficile was isolated by selective cultivation from two dead and a third severely affected elephant. Four isolates were obtained and found positive for toxin A and B by PCR. All isolates were positive in a toxigenic culture assay and toxin was demonstrated in the intestinal content from one of the fatal cases and in a surviving but severely affected elephant. PCR ribotyping demonstrated that the C. difficile isolates shared an identical profile, which was different from an epidemiologically unrelated strain, indicating that the outbreak was caused by the same C. difficile clone. It is speculated that the feeding of large quantities of broccoli, a rich source of sulforaphane, which has been shown to inhibit the growth of many intestinal microorganisms may have triggered a subsequent overgrowth by C. difficile. This is the first report of C. difficile as the main cause of fatal enterocolitis in elephants. The findings emphasize the need to regard this organism as potentially dangerous for elephants and caution is recommended concerning antibiotic treatment and feeding with diets containing antimicrobials, which may trigger an expansion of a C. difficile population in the gut.

Animals↗
PubMed1997

Hospital-acquired Clostridium difficile diarrhoea and herd immunity.

Clostridium difficile diarrhoea represents a significant health-service burden. We recently experienced an outbreak of C difficile diarrhoea associated with increased use of cefotaxime. The question we pose in this paper is how did the introduction and withdrawal of a single antibiotic so greatly affect rates of C difficile diarrhoea? Other antibiotics had nearly as high a risk of causing diarrhoea as cefotaxime, and the majority of patients never received cefotaxime. We believe that such outbreaks of C difficile diarrhoea are best understood in terms of a population model, and that taking antibiotics like cefotaxime should be thought of as a population rather than an individual risk factor. We postulate a herd-immunity model of C difficile diarrhoea, and examine the implications of this hypothesis.

Aged↗
PubMed2002

Clostridium difficile infections in animals with special reference to the horse. A review.

In human medicine, Clostridium (C.) difficile is since many years a well-known cause of nosocomial diarrhea induced by antibiotic treatment. In horses, C. difficile was recently suggested as a possible enteric pathogen. The bacterium is associated with acute colitis in mature horses following treatment with antibiotics. C. difficile, and/or its cytotoxin, is also associated with acute colitis in mares when their foals are being treated with erythromycin and rifampicin for Rhodococcus equi pneumonia. The colitis can have resulted from an accidental ingestion of erythromycin by the mares. In an experimental study it was also demonstrated in mature horses that erythromycin can induce severe colitis associated with proliferation of C. difficile. A new interesting finding was that in healthy foals younger than 14 days, C. difficile was isolated from every third foal whereas older foals proved negative. In this paper the current state of knowledge of C. difficile infections in animals, especially in horses, is reviewed. A short description is given of the historical background of Clostridium difficile and the antibiotic-associated colitis and diarrhea caused by infection with this bacterium. The taxonomy of Clostridium difficile is described extensively. A summary is given of the diseases associated with clostridia infections in animals. Special attention is paid to the pathogenesis, epidemiology, clinical symptoms, laboratory diagnosis, and pathology of Clostridium difficile infections in horses. Finally, some other bacterial causes of colitis in horses are discussed shortly.

Animals↗
PubMed1994

Effectiveness of liquid soap vs. chlorhexidine gluconate for the removal of Clostridium difficile from bare hands and gloved hands.

OBJECTIVE: To compare liquid soap versus 4% chlorhexidine gluconate in 4% alcohol for the decontamination of bare or gloved hands inoculated with an epidemic strain of Clostridium difficile. DESIGN: C difficile (6.7 log10 colony-forming units [CFU], 47% spores), was seeded onto bare or latex gloved hands of ten volunteers and allowed to dry. Half the volunteers initially washed with soap and half with chlorhexidine, followed by the other agent 1 week later. Cultures were done with Rodac plates at three sites on the hand: finger/thumbtips, the palmar surfaces of the fingers, and the palm. Statistical comparison was by paired Student's t test. RESULTS: On bare hands, soap and chlorhexidine did not differ in residual bacterial counts on the finger/thumbtips (log10 CFU, 2.0 and 2.1, P = NS) and fingers (log10 CFU, 2.4 and 2.5, P = NS). Counts were too high on bare palms to quantitate. On gloved hands, soap was more effective than chlorhexidine on fingers (log10 CFU 1.3 and 1.7, P < .01) and palms (log10 CFU 1.5 and 2.0, P < .01), but not finger/thumbtips (log10 CFU 1.6 with each, P = NS). Residual C difficile counts were lower on gloved hands than bare hands (P < 0.01 to < 0.0001). CONCLUSIONS: The two agents did not differ significantly in residual counts of C difficile on bare hands, but on gloved hands residual counts were lower following soap wash than following chlorhexidine wash. These observations support the use of either soap or chlorhexidine as a handwash for removal of C difficile, but efficacy in the prevention of C difficile transmission must be determined by prospective clinical trials.

Aerosols↗
PubMed2000

In vitro and in vivo activities of nitazoxanide against Clostridium difficile.

We have used the hamster model of antibiotic-induced Clostridium difficile intestinal disease to evaluate nitazoxanide (NTZ), a nitrothiazole benzamide antimicrobial agent. The following in vitro and in vivo activities of NTZ in the adult hamster were examined and compared to those of metronidazole and vancomycin: (i) MICs and minimum bactericidal concentrations (MBCs) against C. difficile, (ii) toxicity, (iii) ability to prevent C. difficile-associated ileocecitis, and (iv) propensity to induce C. difficile-associated ileocecitis. The MICs and MBCs of NTZ against 15 toxigenic strains of C. difficile were comparable to those of vancomycin or metronidazole. C. difficile-associated ileocecitis was induced with oral clindamycin and toxigenic C. difficile in a group of 60 hamsters. Subgroups of 10 hamsters were given six daily intragastric treatments of NTZ (15, 7.5, and 3.0 mg/100 g of body weight [gbw]), metronidazole (15 mg/100 gbw), vancomycin (5 mg/100 gbw), or saline (1 ml/100 gbw). Animals receiving saline died 3 days post-C. difficile challenge. During the treatment period, NTZ (>/=7.5 mg/100 gbw), like metronidazole and vancomycin, prevented outward manifestations of clindamycin-induced C. difficile intestinal disease. Six of ten hamsters on a scheduled dose of 3.0 mg of NTZ/100 gbw survived for the complete treatment period. Of these surviving animals, all but three died of C. difficile disease by between 3 and 12 days following discontinuation of antibiotic therapy. Another group of hamsters received six similar daily doses of the three antibiotics, followed by an inoculation with toxigenic C. difficile. All of the NTZ-treated animals survived the 15-day postinfection period. Upon necropsy, all hamsters appeared normal: there were no gross signs of toxicity or C. difficile intestinal disease, nor was C. difficile detected in the cultures of the ceca of these animals. By contrast, vancomycin and metronidazole treatment induced fatal C. difficile intestinal disease in 20 and 70% of recipients, respectively.

Animals↗
PubMed2004

In-vitro activity of nisin against clinical isolates of Clostridium difficile.

Nisin is a cationic peptide produced by Lactococcus lactis. Its activity against clinical isolates of Clostridium difficile was compared to that of vancomycin and metronidazole by minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and time-kill studies. Nisin was more active than the other agents, with a MIC90 of 0.256 mg/L and strong bactericidal activity. Nisin may be a promising agent for the management of C. difficile associated diarrhea.

Anti-Bacterial Agents↗
PubMed2002

[Epidemiology of Clostridium difficile infection].

Pseudomembranous colitis (PMC), antibiotic-associated diarrhoea (AAD), and colitis (AAC) caused by Clostridium difficile are recognized as complications of antibiotic treatment (cephalosporins, penicillins, clindamycin and others). Two groups are particularly at risk: older and immunocompromised patients. In recent years C. difficile has been recognized as a common nosocomial pathogen. To understand the epidemiology of the C. difficile infection, many outbreaks have been investigated by various methods. In the paper we reviewed different methods of C. difficile typing and discussed the epidemiology of C. difficile-associated infections in light of recent publications.

Age Factors↗
PubMed1998

Evaluation of four methods for detection of Clostridium difficile or C. difficile toxin: cytotoxin assay, culture, latex agglutination, and a new rapid immunoassay (C. difficile toxin A test).

The performance of C. difficile toxin A test (Oxoid, Basingstoke, UK), an immunoassay for the detection of C. difficile toxin A in fecal samples, for the diagnosis of C. difficile-associated diarrhea was compared with those of cytotoxin assay, culture, and a latex agglutination assay (Culturette Brand CDT Rapid Clostridium difficile test; Becton Dickinson, Cockeysville, MD). A total of 105 stool specimens from 71 patients were tested. Of the 105 specimens analyzed, 6 (5.7%) samples were positive by all four methods, 66 samples (62.9%) were concordant negative. Two patients fulfilled all clinical criteria for C. difficile-associated diarrhea but had negative results in the cytotoxicity tests. The sensitivity and specificity, as determined against cytotoxin assay results, were, respectively 52.4% and 93.9% for latex agglutination, 68.2% and 84.3% for rapid immunoassay and 36.4% and 92.8% for culture. No single laboratory test yields a definitive diagnosis of C. difficile-associated diarrhea at present. Test results by any of these methods must be used in conjunction with patient history when making the diagnosis to avoid indiscriminate treatment of individuals without disease or the lack of treatment for individuals with a serious infection. We consider the rapid and easy-to-perform C. difficile toxin A test not to be an essential adjunct for the diagnosis of C. difficile-associated diarrhea.

Adolescent↗