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Survey of incidence of Clostridium difficile infection in Canadian hospitals and diagnostic approaches.

A questionnaire relating to Clostridium difficile disease incidence and diagnostic practices was sent to 380 Canadian hospitals (all with > 50 beds). The national questionnaire response rate was 63%. In-house testing was performed in 17.6, 61.5, and 74.2% of the hospitals with < 300, 300 to 500, and > 500 beds, respectively. The average test positivity rates were 17.2, 15.3, and 13.2% for hospitals with < 300, 300 to 500, and > 500 beds, respectively. The average disease incidences were 23.5, 30.8, and 40.3 cases per 100,000 patient days in the hospitals with < 300, 300 to 500, and > 500 beds, respectively. In the 81 hospitals where in-house testing was performed, cytotoxin testing utilizing tissue culture was most common (44.4%), followed by enzyme-linked immunosorbent assay (38.3%), culture for toxigenic C. difficile (32.1%), and latex agglutination (13.6%). The clinical criteria for C. difficile testing were variable, with 85% of hospitals indicating that a test was done automatically if ordered by a doctor. Our results show that C. difficile-associated diarrhea is a major problem in hospitals with > or = 200 beds. Despite a lower disease incidence in smaller hospitals, there was a higher diagnostic test positivity rate. This may reflect the preference of smaller hospitals for culture and latex agglutination tests.

Bacterial Proteins↗

Evaluation of repetitive element sequence-based PCR as a molecular typing method for Clostridium difficile.

Repetitive element sequence-based PCR (rep-PCR) is a typing method that enables the generation of DNA fingerprinting that discriminates bacterial strains. In this study, we evaluated the applicability of rep-PCR in typing Clostridium difficile clinical isolates. The results obtained by rep-PCR were compared with those obtained by pulsed-field gel electrophoresis (PFGE) and PCR ribotyping. A high correspondence between pattern differentiations produced by rep-PCR and PFGE was observed, whereas PCR ribotyping showed a lower level of discriminatory power.

Bacterial Typing Techniques↗

ICD-9 codes and surveillance for Clostridium difficile-associated disease.

We conducted a retrospective cohort study to compare Clostridium difficile-associated disease rates determined by C. difficile-toxin assays and International Classification of Diseases, 9th Revision (ICD-9) codes. The correlation between toxin assay results and ICD-9 codes was good (K = 0.72, p<0.01). The sensitivity of the ICD-9 codes was 78% and the specificity was 99.7%.

Bacterial Toxins↗

[Detection of toxin producing strains of Clostridium difficile using rapid diagnostic methods].

Feces of 53 patients from different hospital wards suffering from long term post-antibiotic therapy diarrhea were tested. For direct detection of C. difficile toxin A, in samples TCD (Becton-Dickinson), and C. difficile Toxin A Test (Oxoid) tests were used. Toxin A was detected in 16 samples (29.6% tested). C. difficile strains were isolated from 40% of the fecal samples. Toxin A was detected in 25 Clostridium difficile strains with commercial tests and toxin B was detected using McCoy cell line. Toxin A was not produced by 3 C. difficile strains in vitro, but toxin B was produced by all strains. The polymerase chain reaction (PCR) test showed that all isolated strains possess genes of toxins A and B.

Adolescent↗

Development and clinical evaluation of an amplified flow cytometric fluoroimmunoassay for Clostridium difficile toxin A.

A rapid (2 h) amplified flow cytometric fluoroimmunoassay (AFCF) for Clostridium difficile toxin A was developed and compared with the cytotoxin assay (CTA) and culture of the organism from stool specimens from patients with suspected C. difficile-associated gastrointestinal disease (CAD). For this assay polyclonal antitoxin A was attached to 10-microns diameter and monoclonal antitoxin A was attached to fluorescent 0.1 micron-diameter polystyrene microspheres. The microspheres and sample were reacted together as in a conventional double-antibody sandwich assay. However, laser flow cytometric measurement allowed the omission of separation and washing steps by gating on light scattered by the larger microspheres and measuring only the fluorescence associated with these particles. The amount of fluorescence from the attached 0.1 micron microspheres was dependent on the concentration of toxin A in the sample. The AFCF detected purified toxin A at levels of 1 pg/ml and was linear from 1 to 40 pg/ml. The AFCF was compared with the CTA and culture of C. difficile for clinical use by comparing results from 198 stool specimens from patients with suspected CAD. The AFCF was 85.7% sensitive and 95.8% specific relative to the CTA, and 85.2% sensitive and 98.3% specific compared to the culture assay. If the isolation of toxigenic C. difficile or the patients clinical course was considered indicative of CAD, the sensitivities of the AFCF, CTA, and culture assay were 77.4%, 67.7% and 96.8%, respectively. The AFCF demonstrated a specificity of 98.8%, while both CTA and culture had a specificity of 100%.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

In vitro effects of Clostridium difficile toxins on hepatocytes.

BACKGROUND: Clostridium difficile infections are associated with development of the systemic inflammatory response, including the production of hepatic acute phase proteins. Lipopolysaccharide (LPS) directly stimulates the production of at least one of these proteins, a 23-kDa acute phase protein (the LPS-induced protein, or LIP) by murine hepatocytes in vitro. The aim of the present study was to determine if C. difficile toxins also stimulated the synthesis of this protein in vitro. METHODS: Cultured murine hepatocytes were treated for 24 h with various concentrations of C. difficile culture extract or purified toxins A and B in the presence or absence of dexamethasone or interleukin-1 (IL-1) receptor antagonist (IL-1 RA). The cells were then metabolically radiolabeled with [35S]methionine. Secretory proteins were identified using electrophoresis and autoradiography, and their synthesis was quantitated by image analysis of the autoradiograms. RESULTS: The C. difficile culture extract, at dilutions as low as 1:200,000, significantly stimulated LIP synthesis in vitro. Toxins A and B, at concentrations as low as 1.6 and 0.02 pg/ml, respectively, also induced production of this protein. Dexamethasone further augmented C. difficile toxin-stimulated synthesis of LIP, but IL-1 RA inhibited the effects of these toxins on the synthesis of this protein. Only minimal quantities of IL-1 were found in culture supernatants following treatment with the toxins. CONCLUSIONS: C. difficile toxins A and B, at very low concentrations, stimulate hepatocyte acute phase protein synthesis. Even though IL-1 RA inhibits this process, it does not appear that local production of IL-1 mediates the action of these toxins.

Acute-Phase Proteins↗

Pseudomembranous colitis associated with chemotherapy with 5-fluorouracil.

Pseudomembranous colitis is frequently associated with antibiotics and more rarely with chemotherapeutic agents such as 5-fluorouracil. The objective of this study is to show that it is possible to confuse this infection with chemotherapy associated toxicity. We present a 54 year old woman who underwent surgery for colorectal cancer and in the first cycle of chemotherapy with 5-fluorouracil developed pseudomembranous colitis. We detected the toxin B of Clostridium difficile in stools and we began early antibiotic treatment. Thus, in patients with post chemotherapy neutropenia and diarrhoea that develop negatively, we have to rule out this infection.

Adenocarcinoma↗

Evaluation of microbicidal activity of a new disinfectant: Sterilox 2500 against Clostridium difficile spores, Helicobacter pylori, vancomycin resistant Enterococcus species, Candida albicans and several Mycobacterium species.

The microbicidal activity of a new disinfectant Sterilox, a super-oxidized water, containing a mixture of oxidizing substances, was tested against Clostridium difficile spores, Helicobacter pylori, vancomycin resistant Enterococcus species, Candida albicans and several Mycobacterium species using membrane filters. All tests were performed in duplicate with and without added horse serum at 1% and 5% v/v. Distilled water, 0.35% peracetic acid (Nu-Cidex) and 2% glutaraldehyde were included as controls. Sterilox: spore suspension (9:1 v/v) achieved log10 kill of > 5 with 5% horse serum in 2 min against H. pylori, vancomycin resistant Enterococcus species, C. albicans and four atypical Mycobacterium species: M. avium, M. chelonei, M. xenopi and M. smegmatis. Sporicidal activity of Sterilox against Clostridium difficile was markedly diminished in the presence of 5% horse serum. Sterilox may be an effective alternative in endoscopy units, as it is a potent microbicidal agent and the manufacturer claims it is not corrosive to metal and is nontoxic to biological tissues.

Anti-Bacterial Agents↗

Microbiological studies of the enterocolitis of Hirschsprung's disease.

The results of a prospective study of 20 cases of newly diagnosed Hirschsprung's disease (nine of whom developed enterocolitis) and 10 normal controls showed no variations in the bacterial flora (including Clostridium difficile) in the stools of the groups studied. Viral studies showed that rotavirus was present in the stools of seven of the nine cases of enterocolitis during the episode. We suggest that Hirschsprung's enterocolitis may have a complex infective aetiology and that rotavirus plays a part.

Bacteria↗

Clostridium difficile-associated diarrhea in adults.

Clostridium difficile is the most important cause of nosocomial diarrhea in adults. Illness may range from mild watery diarrhea to life-threatening colitis. An antecedent disruption of the normal colonic flora followed by exposure to a toxigenic strain of C. difficile are necessary first steps in the pathogenesis of disease. Diagnosis is based primarily on the detection of C. difficile toxin A or toxin B. First-line treatment is with oral metronidazole therapy. Treatment with oral vancomycin therapy should be reserved for patients who have contraindications or intolerance to metronidazole or who fail to respond to first-line therapy.

Anti-Infective Agents↗

Extraintestinal Clostridium difficile: 10 years' experience at a tertiary-care hospital.

OBJECTIVE: To determine the clinical characteristics of patients with extraintestinal Clostridium difficile (ECD). MATERIAL AND METHODS: All cultures obtained during a 10.5-year period (from Jan. 1, 1985, to Jun. 30, 1995) at a tertiary-care hospital were retrospectively examined. The medical records of patients from whom ECD was isolated were then reviewed. RESULTS: Fourteen patients from whom ECD was cultured were identified. Thirteen of these patients (93%) had underlying systemic disease. All but one patient had recent exposure to antibiotics, and all had major bowel pathologic conditions. Nine patients had colon perforation. Of the eight patients in whom the colonic mucosa was directly inspected at operation or endoscopy, only two had evidence of pseudomembranous colitis. Five patients (36%) had documentation of recent diarrhea. ECD was isolated from intraperitoneal sites (in nine patients), blood cultures (in three), a perianal abscess, and a prosthetic hip joint. In 13 patients (93%), the infection was polymicrobial. Seven of the 13 inpatients (54%) survived to dismissal. CONCLUSION: C. difficile is a rare isolate outside of the gastrointestinal tract. ECD is found in patients with systemic illness who have been hospitalized (usually for an extended period), have intestinal pathologic conditions, and have received antibiotics. The isolation of ECD portends a poor prognosis.

Abscess↗

Clostridium difficile pilot study: effects of probiotic supplementation on the incidence of C. difficile diarrhoea.

Colonic infection with Clostridium difficile, leading to pseudomembranous colitis, is a common complication of antibiotic therapy, especially in elderly patients. It has been suggested that non-pathogenic probiotic bacteria might prevent the development and recurrence of C. difficile infection. This double-blind, placebo-controlled study examines the role of probiotic administration in the prevention of C. difficile-associated diarrhoea (CDAD) in elderly patients receiving antibiotic therapy. Consecutive patients (150) receiving antibiotic therapy were randomised to receive either a probiotic containing both Lactobacillus and Bifidobacterium or placebo for 20 days. Upon admission to hospital, bowel habit was recorded and a faecal sample taken. Trial probiotic or placebo was taken within 72 h of prescription of antibiotics, and a second stool sample was taken in the event of development of diarrhoea during hospitalisation or after discharge. Of the randomised patients, 138 completed the study, 69 with probiotics in conjunction with antibiotics and 69 with antibiotics alone. On the basis of development of diarrhoea, the incidence of samples positive for C. difficile-associated toxins was 2.9% in the probiotic group compared with 7.25% in the placebo-control group. When samples from all patients were tested (rather than just those developing diarrhoea) 46% of probiotic patients were toxin-positive compared with 78% of the placebo group.

Clostridioides difficile↗

Clostridium difficile-associated diarrhoea in patients with human immunodeficiency virus infection: a case-control study.

OBJECTIVE: To evaluate the prevalence of, risk factors for, treatment and outcome of Clostridium difficile-associated diarrhoea (CDAD) in patients with human immunodeficiency virus (HIV) infection. DESIGN: A prospective case-control study, conducted between January 1992 and April 1994. SETTING: Department of Infectious Diseases in a large university hospital with HIV in- and out-patient units. PATIENTS AND METHODS: The study included 124 patients grouped as follows: 31 HIV-infected patients with CDAD (group A); 31 HIV-seronegative patients with CDAD (group B) and 62 HIV-infected patients without CDAD (group C). The patients in group B and C were selected randomly during the study period. RESULTS: The prevalence of CDAD in HIV-infected patients was 3.1% compared with 1.6% in HIV-seronegative patients (P = 0.02). On univariate analysis, the predisposing factors in group A were antibiotic use in the 4 weeks prior to the onset of CDAD (P = 0.03 versus group C), prolonged hospitalization (over 20 days; P = 0.04), low levels of circulating CD4+ cells (P = 0.03) and use of antacids (P = 0.04). The antibiotics significantly associated with CDAD were trimethoprim-sulfamethoxazole (P = 0.02 versus group C), third generation cephalosporins (P = 0.03) and clindamycin (P = 0.03). On multivariate analysis of the risk factors, the use of antibiotics was the sole independent risk factor for CDAD (P = 0.03). The clinical symptoms of CDAD were more severe in HIV-infected patients than in controls. Three patients in group A (9.7%) had one relapse and one patient (3.2%) experienced chronic diarrhoea. The outcome of CDAD was not influenced by the number of circulating polymorphonuclear cells and CD4+ cells. No difference in the survival curves of AIDS patients with or without CDAD, stratified according to age, sex and CD4+ cell count was observed. CONCLUSIONS: Our data suggest that CDAD is more common in HIV-infected patients, particularly those receiving antibiotic therapy, than in HIV-seronegative patients. Since C. difficile can cause severe and recurrent or chronic infections in HIV-infected patients, CDAD must be always considered in the differential diagnosis of diarrhoea in patients with AIDS and AIDS-related conditions.

AIDS-Related Opportunistic Infections↗

Detection of Clostridium difficile toxins in stools. Comparison between a new enzyme immunoassay for toxin A and other routine tests.

An enzyme-linked immunosorbent assay (ELISA) for detection of Clostridium difficile toxin A (enterotoxin) using a monoclonal antibody is described. No cross-reaction was observed with any of the Clostridium species tested except for toxigenic Clostridium difficile. One hundred and eight stool specimens from hospitalized patients harbouring C. difficile in their intestine and 43 samples negative for C. difficile isolation were studied to compare this test with a cytotoxicity assay, the isolation of toxigenic C. difficile and the commercial EIA Premier test (Meridian). As compared with the cytotoxicity assay and EIA, specificity was 91 and 100%, while sensitivity 83 and 74% respectively. This ELISA technique could be used as a routine test for toxin A detection in stools.

Adolescent↗

[Diarrhea associated with Clostridium difficile: one-year experience in a general hospital].

Clostridium difficile is considered the most common cause of nosocomial acquired diarrhoea, with frequencies differing widely from one institution to another. So far, it is a scarcely reported condition in Spain. In the present study 129 episodes of Clostridium difficile associated diarrhoea (CDAD) occurred in 120 patients in a 2,000-bed hospital in 1994 is reported. All cases were diagnosed by demonstrating cytotoxicity on cellular lines (MRC-5) from feces or from the strain isolated from a culture medium (CCFA). The overall incidence was 2.4 episodes every 1,000 admissions. Twenty-eight out of the 120 patients (23%) were HIV-positive patients, that is, an incidence of 30 episodes every 1,000 admissions. No significant differences were observed regarding the presentation and clinical course between HIV-positive and HIV-negative patients, with the exception of the antimicrobial agents used previously. Forty-two percent of patients had undergone surgery and 97% had received antimicrobials in the 8 weeks before the CDAD episode, with an average of 3.3 antibiotics per patient. Out of the 129 episodes, 72.8% were treated correctly. A total of 11.7% of patients responded exclusively to the discontinuation of the antimicrobials that were being administered. Eighty-three patients were treated with specific antibiotics, 59 with oral vancomycin, and 24 with metronidazole. Seventy-six patients (91.5%) responded to the initial therapy, 5 relapsed (6%), and 2 (2.5%) failed. The associated mortality rate was 0.7%. C. difficile can be a relevant cause of nosocomial diarrhoea in our setting, particularly in HIV-positive patients, but also in other patients. Its early diagnosis and appropriate therapy can contribute to decrease a relevant cause of morbidity in inpatients.

Anti-Bacterial Agents↗

Surgical aspects of Clostridium difficile colitis.

BACKGROUND: There has been a marked increase in the number of surgical patients developing Clostridium difficile colitis. The epidemiology, pathogenesis, diagnosis and management of C. difficile infection were reviewed from a surgical perspective. METHODS: A literature review was carried out based primarily on a Medline search of all English language publications containing the term C. difficile. RESULTS: The recent dramatic increase in diagnosis of C. difficile infection amongst surgical patients results from heightened awareness of the condition, better methods of diagnosis, more widespread use of antibiotics for treatment and prophylaxis, and the increasing numbers of elderly and immunocompromised patients with malignancy, sepsis, and (multiple) organ failure being cared for within intensive therapy and high-dependency units. In addition to morbidity and mortality, the economic burden of C. difficile infection in terms of delayed discharge and other hospital costs is considerable. CONCLUSION: Appropriate use of antibiotics, isolation of affected patients and meticulous hygiene measures on the part of staff are vital if the morbidity, mortality and economic consequences of this nosocomial infection are to be minimized.

Antibiotic Prophylaxis↗

Gastrointestinal toxicity and Clostridium difficile diarrhea in patients treated with paclitaxel-containing chemotherapy regimens.

OBJECTIVE: The objective of this study was to determine the incidence of grade 3 and 4 gastrointestinal toxicity and the prevalence of Clostridium difficile-associated diarrhea (CDAD) in patients with gynecologic malignancies treated with paclitaxel-based chemotherapy regimens. METHODS: We retrospectively reviewed the medical records of all patients treated on the Gynecology Service at Memorial Sloan-Kettering Cancer Center from January 1, 1993 to July 1, 1996. We identified all patients treated with paclitaxel during this period and determined the total number of patients hospitalized for symptoms of gastrointestinal toxicity, including nausea, vomiting, diarrhea, and dehydration, within 4 weeks of chemotherapy, as well as the incidence of CDAD among these patients. RESULTS: Six hundred and twenty-four patients were treated with paclitaxel-containing chemotherapy regimens during the study period, including 55 patients who were treated on a "dose-dense" high-dose protocol for advanced ovarian cancer. Among these, 149 patients (24%) were hospitalized for symptoms of gastrointestinal toxicity. During the study period, a total of 40 cases of CDAD were reported among hospitalized patients on the Gynecology Service and 24 (60%) of these cases occurred in patients who had received paclitaxel within the prior 4 weeks. CONCLUSIONS: The occurrence of CDAD in patients receiving paclitaxel-containing chemotherapy is not rare and can result in severe dehydration requiring hospitalization. The risk of C. difficile colitis appears to be 2.2% in patients receiving standard-dose regimens and as high as 20% in patients receiving high-dose regimens. This etiology should be considered and treated early in patients presenting with symptoms of gastrointestinal toxicity subsequent to chemotherapy treatments.

Anti-Bacterial Agents↗

Pseudomembranous colitis with associated fulminant ileitis in the defunctionalized limb of a jejunal-ileal bypass. Report of a case.

Presented is what is believed to be the first reported case of a defunctionalized limb of small intestine serving as a reservoir for Clostridium difficile. Because of the altered intestinal continuity, the ensuing enteritis and colitis failed to respond to nonoperative management. Current treatment strategies are reviewed. Surgical intervention, including restoration of normal gastrointestinal continuity, should be considered early in the hospital course of this patient population.

Aged↗