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PubMed2003

Pseudomembranous colitis caused by toxin A-negative/toxin B-positive variant strain of Clostridium difficile.

We report the first case of pseudomembranous colitis (PMC) due to a toxin A-negative, toxin B-positive strain (toxin A variant strain) of Clostridium difficile in Japan. The toxin A variant strain of C. difficile is capable of causing PMC and is undetectable in clinical laboratories that use only toxin A immunoassays for C. difficile testing. If C. difficile-associated diarrhea is clinically suspected and toxin A is not detected, then the possibility of a toxin A variant strain should be considered, and further diagnostic testing, such as polymerase chain reaction (PCR) analysis for the detection of C. difficile toxin genes should be performed.

Aged↗
PubMed1991

Control of nosocomial transmission of Clostridium difficile based on sporadic case surveillance.

The recognition of a cluster of antibiotic-associated nosocomial Clostridium difficile disease (NCDD) caused by serotype C in a surgical ward led to a hospital-wide NCDD surveillance and control program. The initial step included: (a) gas-liquid chromatography screening of inpatients' diarrheal stools; (b) enteric isolation precautions, cohorting and terminal room disinfection in wards with a cluster of two or more NCDD cases per month. During a 12-month period, the quarterly incidence of NCDD remained unchanged and six new clusters of serotype C, K, and H infections occurred, giving a global incidence of 1.5/1,000 admissions. C. difficile spores were recovered from 36.7% surfaces of case patient rooms versus 6.7% in control rooms. More intensive control measures were evaluated: (a) culture screening of inpatients' diarrheal stools; (b) early therapy, enteric isolation precautions, and daily meticulous room disinfection for each sporadic NCDD case. Surface disinfection reduced the contamination level four-fold (p = 0.04). In the following 12 months, no cluster occurred and the incidence of NCDD fell to 0.3/1,000 admission (protective efficacy 73%, 95% confidence interval: 46-87%). These observations suggest that early therapy, isolation precautions, and surface disinfection, focused on patients with sporadic NCDD detected by active surveillance, can prevent nosocomial transmission of C. difficile.

Adolescent↗
PubMedDate not supplied

Toxin production by an emerging strain of Clostridium difficile associated with outbreaks of severe disease in North America and Europe.

BACKGROUND: Toxins A and B are the primary virulence factors of Clostridium difficile. Since 2002, an epidemic of C difficile-associated disease with increased morbidity and mortality has been present in Quebec province, Canada. We characterised the dominant strain of this epidemic to determine whether it produces higher amounts of toxins A and B than those produced by non-epidemic strains. METHODS: We obtained isolates from 124 patients from Centre Hospitalier Universitaire de Sherbrooke in Quebec. Additional isolates from the USA, Canada, and the UK were included to increase the genetic diversity of the toxinotypes tested. Isolate characterisation included toxinotyping, pulsed-field gel electrophoresis (PFGE), PCR ribotyping, detection of a binary toxin gene, and detection of deletions in a putative negative regulator for toxins A and B (tcdC). By use of an enzyme-linked immunoassay, we measured the in-vitro production of toxins A and B by epidemic strain and non-dominant strain isolates. FINDINGS: The epidemic strain was characterised as toxinotype III, North American PFGE type 1, and PCR-ribotype 027 (NAP1/027). This strain carried the binary toxin gene cdtB and an 18-bp deletion in tcdC. We isolated this strain from 72 patients with C difficile-associated disease (58 [67%] of 86 with health-care-associated disease; 14 [37%] of 38 with community-acquired disease). Peak median (IQR) toxin A and toxin B concentrations produced in vitro by NAP1/027 were 16 and 23 times higher, respectively, than those measured in isolates representing 12 different PFGE types, known as toxinotype 0 (toxin A, median 848 microg/L [IQR 504-1022] vs 54 microg/L [23-203]; toxin B, 180 microg/L [137-210] vs 8 microg/L [5-25]; p<0.0001 for both toxins). INTERPRETATION: The severity of C difficile-associated disease caused by NAP1/027 could result from hyperproduction of toxins A and B. Dissemination of this strain in North America and Europe could lead to important changes in the epidemiology of C difficile-associated disease.

Bacterial Proteins↗
PubMed2003

Clostridium difficile colitis associated with hemolytic-uremic syndrome.

The authors report the case of a 46-year-old woman who presented with vomiting and profuse bloody diarrhea. Laboratory studies were significant for a hematocrit of 27% and lactate dehydrogenase of 5,394 U/L (5,394 U/L). Her renal function deteriorated rapidly with a peak creatinine of 12.4 mg/dL (1,096.4 micromol/L), and platelet count dropped simultaneously to a nadir of 123,000/microL (123 x 10(9)/L]. Schistocytes were observed in peripheral blood smear. Stool was positive for Clostridium difficile toxin A by enzyme immunoassay (EIA). Stool assay for Shiga-like toxin was negative by EIA, and stool cultures returned negative for Escherichia coli O157:H7 and other enteric pathogens. A diagnosis of C difficile colitis associated with hemolytic-uremic syndrome was made; the patient received plasmapheresis and recovered with no relapse after 10 months of follow-up. This is the second reported case of C difficile colitis associated with hemolytic-uremic syndrome in an adult.

Clostridioides difficile↗
PubMed2005

Treatment of Clostridium difficile-associated disease: old therapies and new strategies.

Clostridium difficile-associated disease (CDAD) causes substantial morbidity and mortality. The pathogenesis is multifactorial, involving altered bowel flora, production of toxins, and impaired host immunity, often in a nosocomial setting. Current guidelines recommend treatment with metronidazole; vancomycin is a second-line agent because of its potential effect on the hospital environment. We present the data that led to these recommendations and explore other therapeutic options, including antimicrobials, antibody to toxin A, probiotics, and vaccines. Treatment of CDAD has increasingly been associated with failure and recurrence. Recurrent disease may reflect relapse of infection due to the original infecting organism or infection by a new strain. Poor antibody responses to C difficile toxins have a permissive role in recurrent infection. Hospital infection control and pertinent use of antibiotics can limit the spread of CDAD. A vaccine directed against C difficile toxin may eventually offer a solution to the CDAD problem.

Anti-Infective Agents↗
PubMed2002

Clostridium difficile, atopy and wheeze during the first year of life.

Differences have been suggested to occur in the composition of intestinal microflora from allergic and non-allergic children. In this study we used a semi-quantitative enzyme-linked immunosorbent assay (ELISA) for the measurement of Clostridium difficile-specific immunoglobulin G (IgG) (CDIgG). CDIgG was excellent in differentiating between adults with or without Cl. difficile colitis (absorbance levels, positive vs. negative controls: geometric mean (GM) 0.301, 95% CI: 0.289-0.314 vs. GM 0.167, 95% CI: 0.155-0.181; mean difference 1.8-fold, 95% CI: 1.65-1.95; p < 0.0001). We used this technique to investigate whether there are any differences between atopic wheezy infants and non-atopic non-wheezy controls. In a prospective cohort study (n = 390) 10 patients were identified at 1 year of age (atopic, history of recurrent wheeze) and matched (gender, month of birth, exposure to Der p 1, Fel d 1 and Can f 1) with a control group of infants (non-atopic, no history of wheeze). The patients had significantly higher Cl. difficile-specific IgG absorbance levels (GM 0.298, 95% CI: 0.249-0.358) compared with controls (GM 0.235, 95% CI: 0.201-0.274; mean difference 1.27-fold, 95% CI: 1.07-1.50; p = 0.01). These results suggest that there may be differences in the composition of intestinal microflora between allergic and non-allergic infants at 1 year of age, with allergic children having higher Cl. difficile IgG antibody levels.

Antibody Specificity↗
PubMed2001

Molecular analysis of relapse vs re-infection in HIV-positive patients suffering from recurrent Clostridium difficile associated diarrhoea.

Recurrence is a major complication of Clostridium difficile associated diarrhoea, especially in human immunodeficiency virus (HIV) positive patients, and it is important to distinguish between relapse and re-infection in recurrent episodes. The aim of our study was to analyse C. difficile isolates obtained from HIV-positive patients with recurrent diarrhoea in order to distinguish between relapse and re-infection. This analysis was based on the study of DNA similarities among isolates obtained from different episodes within each patient. Relapses occurred in 64% of patients, 32% suffered re-infections and a combination of relapse plus re-infection was seen in 4%. DNA typing methods can be useful tools to characterize recurrent episodes of C. difficile associated disease.

AIDS-Related Opportunistic Infections↗
PubMed1995

Role of Clostridium difficile in childhood diarrhea.

To investigate the etiologic role of Clostridium difficile in childhood acute diarrhea, stool specimens from 618 children with diarrhea and 135 controls without enteric symptoms were examined by cell culture assay for the presence of free toxin B. This toxin was found in 4.2% of the fecal specimens examined without finding a significant difference between cases and controls, suggesting no causal relationship between diarrhea and the presence of free C. difficile toxin B. C. difficile strains isolated from toxin B-positive specimens were characterized by cytotoxin and enterotoxin production and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of EDTA-extracted proteins. All but two isolates produced toxin B and toxin A and the remaining were negative for both toxins. Polyacrylamide gel electrophoresis analysis showed eight electrophoretic types, none of them was clearly related with the cases of diarrhea. The majority of isolates from children with diarrhea did not belong to types previously observed in adults with pseudomembranous colitis or antibiotic-associated diarrhea. This study provides additional evidence that C. difficile is not involved in the etiology of childhood diarrhea.

Bacterial Toxins↗
PubMed2000

Clostridium difficile and vancomycin-resistant enterococcus: the new nosocomial alliance.

OBJECTIVES: The aims of this study were to determine the frequency of the association between Clostridium difficile (C. difficile) and vancomycin-resistant Enterococcus (VRE) and delineate the role of C. difficile coinfection as a predictor of VRE infection versus colonization and adverse outcome. METHODS: Patients with both C. difficile colitis and VRE (CD/VRE) were compared to patients with VRE alone with regard to demographics, comorbidity, prior antibiotic therapy, and coinfection with methicillin-resistant Staphylococcus aureus and funguria. C. difficile as a predictor of VRE infection (VRE-I) versus colonization (VRE-C) and adverse outcome was also studied. RESULTS: Eighty-nine patients with VRE infection or colonization were studied. This included 31 cases of VRE-I and 58 VRE-C. C. difficile was isolated in 17 (19.1%) of patients; of these C. difficile was isolated before VRE in 9 patients and after VRE in 8. The two groups did not differ in age, residence, or comorbidity. C. difficile coinfection was not predictive of VRE-I versus VRE-C, nor was it associated with increased length of stay or mortality. However, the mortality rates in both groups was high, around 30%. A significant association was noted between the use of vancomycin and metronidazole (before the isolation of VRE) and C. difficile coinfection (p = 0.03 and p = 0.001, respectively). A high incidence of nosocomial coinfection with methicillin-resistant Staphylococcus aureus, funguria, and gram-negative sepsis was noted in both groups; the association with funguria was statistically significant (p = 0.029). CONCLUSIONS: In conclusion, C. difficile coinfection is common in patients with VRE infection or colonization and is significantly associated with other nosocomial dilemmas like funguria. This may result in the emergence of highly virulent pathogens including vancomycin-resistant C. difficile, posing new challenges in the management of nosocomial diarrheas.

Aged↗
PubMed1994

Haemagglutination activity of toxigenic and non-toxigenic strains of Clostridium difficile.

Cell extract of Clostridium difficile strains was fractionated by ammonium sulfate precipitation and sulfated cellulofine column chromatography to detect haemagglutination (HA) activity. HA activity without cytotoxicity was detected in fractions eluted at 0.79-0.91 M NaCl in sulfated cellulofine column chromatography of the cell extract in both toxigenic strain VPI 10463 and non-toxigenic strain KZ 1678, while toxin A was detected in fractions eluted at 0.27-0.29 M NaCl. Antisera were prepared with HA substance-containing fractions of the chromatography. Antiserum to the HA substance(s) of strain VPI 10463 neutralised the HA activity of the fractions of strains VPI 10463 and KZ 1678 at nearly the same titres. Antiserum to the HA substance(s) of strain KZ 1678 also neutralised the HA activity of both strains at nearly the same titres as above. These findings suggest that haemagglutinin(s) is commonly produced by C. difficile strains irrespective of toxin A-producing ability.

Animals↗
PubMed2005

Interaction of bismuth subsalicylate with fruit juices, ascorbic acid, and thiol-containing substrates to produce soluble bismuth products active against Clostridium difficile.

Bismuth subsalicylate (BSS), the active ingredient of Pepto-Bismol, has been used for many years to treat various disorders of the gastrointestinal tract. Using mass spectrometry and the agar dilution method, we determined that insoluble BSS interacts with certain dietary components and organic substrates to produce water-soluble products with activity against Clostridium difficile.

Anti-Bacterial Agents↗
PubMed2004

Clostridium difficile associated diarrhoea in hospitalised patients: onset in the community and hospital and role of flexible sigmoidoscopy.

OBJECTIVES: Clostridium difficile associated diarrhoea (CDAD) is a hospital acquired infection in which optimal methods for diagnosis and the scale of the problem in the community remain to be determined. In hospitalised patients with CDAD, we aimed to (i) study patients in whom the onset of diarrhoea was in the community and (ii) investigate the role of bedside flexible sigmoidoscopy in diagnosis. METHODS: Patients with CDAD (onset in hospital or community) were studied prospectively. In those with diarrhoea of unknown aetiology, flexible sigmoidoscopy was compared with stool assay for C difficile cytotoxin. RESULTS: Of 136 patients with CDAD (which was associated with antibiotic exposure in 96%), diarrhoea started in the community in 38 (28%; majority in own home) and while an inpatient in 98 (72%). The majority with CDAD onset in the community had been hospitalised over the preceding 12 months (86.8% v 57.1% in the hospital onset group; p<0.001). In 56 patients with pseudomembranous colitis at sigmoidoscopy, the stool C difficile cytotoxin test was negative in 29 (52%) but toxigenic C difficile was isolated from all of nine stool samples cultured. Of patients with pseudomembranous colitis, 30.4% relapsed over the subsequent 57.7(4.2) days. CONCLUSIONS: In a significant proportion of hospitalised patients with CDAD, diarrhoea started in the community. However, the majority of these had been hospital inpatients previously when they may have acquired C difficile, with the subsequent onset of diarrhoea in the community following exposure to antibiotics. Flexible sigmoidoscopy is superior to the stool C difficile cytotoxin test in a subgroup of patients with pseudomembranous colitis. Sigmoidoscopy should therefore be considered in all hospitalised patients with diarrhoea in whom the stool test for C difficile cytotoxin and enteric pathogens is negative.

Aged↗
PubMed1992

Simultaneous detection of toxin A and toxin B genetic determinants of Clostridium difficile using the multiplex polymerase chain reaction.

A multiplex polymerase chain reaction was developed to simultaneously detect the presence of toxin A and toxin B genes of Clostridium difficile. A 1050-bp fragment of the toxin B gene and a 1217-bp fragment of the toxin A gene were amplified from 42 toxic strains of C. difficile; however, from 10 nontoxic strains the toxin gene fragments were not amplified; these data demonstrate that this multiplex polymerase chain reaction procedure can be used to differentiate between toxic and nontoxic strains. This sensitive and specific multiplex polymerase chain reaction for C. difficile toxins may prove to be a valuable diagnostic procedure.

Bacterial Proteins↗
PubMed2004

Risk of Clostridium difficile diarrhea among hospital inpatients prescribed proton pump inhibitors: cohort and case-control studies.

BACKGROUND: Antibiotic disruption of the normal intestinal flora is a well-known risk factor for Clostridium difficile-associated diarrhea. Reduced gastric acidity has been suggested as a risk factor, and we hypothesized that proton pump inhibitors, because of their potency, may be an independent risk factor for this problem. METHODS: For the cohort study we identified from a pharmacy database 1187 inpatients at a Montreal teaching hospital who received antibiotics over a 9-month period beginning in August 2002. We compared patients in this group who had also received a proton pump inhibitor or an H(2) blocker with patients who had not received acid suppressive therapy. Hospital laboratory reports of positive assay results for C. difficile toxin were used to ascertain cases in the cohort. To assess the possibility that proton pump inhibitors were prescribed to patients who were sicker and had other risk factors for C. difficile infection, we did a case-control study at a second Montreal teaching hospital. Cases were defined as patients who were positive for C. difficile toxin and who had a history of diarrhea (n = 94). Control subjects were selected from among patients who had received an antibiotic and were matched to cases by ward, age within 5 years and class of antibiotics (n = 94). RESULTS: In the cohort study, C. difficile diarrhea developed in 81 (6.8%) of the 1187 patients who received antibiotics while in hospital. In a multivariate analysis, C. difficile diarrhea was significantly associated with use of proton pump inhibitors (adjusted odds ratio [OR] 2.1, 95% confidence interval [CI] 1.2- 3.5), receipt of 3 or more antibiotics (OR 2.1, 95% CI 1.3- 3.4) and admission to a medical ward (OR 4.1, 95% CI 2.3- 7.3). In the case-control study C. difficile diarrhea was associated with female sex (adjusted OR 2.1, 95% CI 1.1-4.0), prior renal failure (adjusted OR 4.3, 95% CI 1.5-11.9), hospital admission in the 3 months before the index admission (adjusted OR 2.6, 95% CI 1.4-5.2) and use of proton pump inhibitors (adjusted OR 2.7, 95% CI 1.4-5.2). INTERPRETATION: Patients in hospital who received proton pump inhibitors were at increased risk of C. difficile diarrhea.

Aged↗
PubMed2000

[Detection of toxin-producing pathogenic bacterial strains by polymerase chain reaction].

Polymerase chain reaction (PCR) was used for detection of pathogenic Clostridium botulinum, Clostridium perfringens, Clostridium difficile, and Escherichia coli. With this aim in view, primers to botulinic toxins types A, B, C1, D, E, F, and G, perfringens enterotoxin, difficile toxin, and types 1 and 2 Shigella-like toxins were chosen and synthesized. Optimal amplification conditions were selected for each pair of primers, with DNA and the respective agent as the reaction mixture matrices. PCR was highly specific and sensitive in all cases. Its sensitivity was 10-100 cells/sample. Among the tested C. botulinum, C. perfringens, C. difficile, and E. coli strains, specific amplification products of expected size were observed only in the strains containing the respective toxin genes. These findings recommend the use of these methods in clinical microbiology. Strains containing type 2 Shigella-like toxin gene were detected among E. coli strains isolated from patients with the hemolytic uremic syndrome, which for the first time indicates that the problem with E. coli epidemic strain O157 is valid for Russia. As a result of our studies, test systems for detection of types A, B, C, D, E, F, and G C. botulinum strains, C. perfringens and C. difficile, and E. coli O157 strains are now available.

Bacterial Toxins↗
PubMed1992

Evaluation of the latex agglutination test for detection of Clostridium difficile.

We compared two Clostridium difficile latex agglutination tests, Meritec from Meridian Diagnostic (Cincinnati, Ohio) and CDT from Becton-Dickinson (Cockeysville, Md), on 289 specimens submitted for tissue culture cytotoxicity using MRC-5 cells. When compared with CDT, the Meritec latex agglutination test had a sensitivity of 90% (26/29), a specificity of 97% (251/260), and a correlation of 96%. Meritec was compared with tissue culture cytotoxicity on 357 specimens. Meritec had a sensitivity of 77% (30/39), a specificity of 93% (298/318), and a correlation of 92%. Clinical review of 10 Meritec +/- tissue culture cytotoxicity minus patients revealed one likely, two probable, and seven doubtful cases of C difficile disease. In contrast, review of 10 Meritec +/- tissue culture cytotoxicity plus patients showed seven likely and three probable cases of C difficile disease. The Meritec is comparable with the CDT latex agglutination test, but is not nearly as sensitive as either tissue culture assay or culture for detection of C difficile disease. A positive latex agglutination test should be confirmed by a tissue culture cytotoxicity assay.

Clostridioides difficile↗