scholarly journals Characterization of clinical Clostridium difficile isolates by PCR ribotyping and detection of toxin genes in Austria, 2006–2007

2008 ◽  
Vol 57 (6) ◽  
pp. 702-708 ◽  
Author(s):  
A. Indra ◽  
D. Schmid ◽  
S. Huhulescu ◽  
M. Hell ◽  
R. Gattringer ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Pseudomembranous Colitis ◽  
Antimicrobial Agents ◽  
Toxic Megacolon ◽  
Health Care Facilities ◽  
Reference Laboratory ◽  
Specimen Collection ◽  
Pcr Ribotyping ◽  
High Level

In order to assess the lethality of Clostridium difficile-associated disease (CDAD) and the PCR ribotypes prevalent in Austria, the Austrian Agency for Health and Food Safety requested isolates of C. difficile from patients in a structured but arbitrary sampling scheme. In the allocated period from February 2006 to January 2007, local hospital laboratories within each of the nine provinces were asked to submit C. difficile isolates from at least ten cases of CDAD. Confirmation of species identification, toxin detection, susceptibility testing against four antimicrobial agents and typing using a PCR ribotyping method were performed at the reference laboratory. In total, 149 isolates of putative C. difficile were submitted, from which 142 were included for study. Antimicrobial susceptibility patterns revealed resistance to clindamycin in 57 % and high-level resistance to moxifloxacin in 38 % of isolates tested. CDAD manifested as diarrhoea (including eight cases of bloody diarrhoea) in 126 cases (88.7 %), as pseudomembranous colitis in 15 cases (10.6 %) and as toxic megacolon in one case. Twelve of the 142 patients died within 30 days of specimen collection (8.45 % lethality). A lethal outcome occurred in 2/15 cases (13.3 %) when pseudomembranous colitis was present and in 10/126 cases (7.9 %) in the absence of pseudomembranous colitis or toxic megacolon. Among the 142 isolates from 25 health-care facilities, 41 PCR ribotype patterns were found. The most frequent ribotypes were AI-5 (including six lethal cases out of 26 patients), 014 (two out of 24) and 053 (one out of 24). The typing patterns demonstrated the occurrence of clusters in hospitals.

Recommendations for Surveillance of Clostridium difficile–Associated Disease

2007 ◽  
Vol 28 (2) ◽  
pp. 140-145 ◽  
Author(s):  
L. Clifford McDonald ◽  
Bruno Coignard ◽  
Erik Dubberke ◽  
Xiaoyan Song ◽  
Teresa Horan ◽  
...  
Keyword(s):  
Intensive Care Unit ◽  
Clostridium Difficile ◽  
Pseudomembranous Colitis ◽  
Ad Hoc ◽  
Toxic Megacolon ◽  
Healthcare Facility ◽  
Case Patient ◽  
Healthcare Settings ◽  
Community Onset ◽  
Laboratory Assay

Background.The epidemiology of Clostridium difficile-associated disease (CDAD) is changing, with evidence of increased incidence and severity. However, the understanding of the magnitude of and reasons for this change is currently hampered by the lack of standardized surveillance methods.Objective and Methods.An ad hoc C. difficile surveillance working group was formed to develop interim surveillance definitions and recommendations based on existing literature and expert opinion that can help to improve CDAD surveillance and prevention efforts.Definitions and Recommendations.A CDAD case patient was defined as a patient with symptoms of diarrhea or toxic megacolon combined with a positive result of a laboratory assay and/or endoscopic or histopathologic evidence of pseudomembranous colitis. Recurrent CDAD was defined as repeated episodes within 8 weeks of each other. Severe CDAD was defined by CDAD-associated admission to an intensive care unit, colectomy, or death within 30 days after onset. Case patients were categorized by the setting in which C. difficile was likely acquired, to account for recent evidence that suggests that healthcare facility-associated CDAD may have its onset in the community up to 4 weeks after discharge. Tracking of healthcare facility–onset, healthcare facility–associated CDAD is the minimum surveillance required for healthcare settings; tracking of community–onset, healthcare facility–associated CDAD should be performed only in conjunction with tracking of healthcare facility–onset, healthcare facility–associated CDAD. Community–associated CDAD was defined by symptom onset more than 12 weeks after the last discharge from a healthcare facility. Rates of both healthcare facility–onset, healthcare facility–associated CDAD and community–onset, healthcare facility–associated CDAD should be expressed as case patients per 10,000 patient–days; rates of community-associated CDAD should be expressed as case patients per 100,000 person-years.

scholarly journals Prevalence and association of PCR ribotypes of Clostridium difficile isolated from symptomatic patients from Warsaw with macrolide-lincosamide-streptogramin B (MLSB) type resistance

2006 ◽  
Vol 55 (2) ◽  
pp. 207-213 ◽  
Author(s):  
Hanna Pituch ◽  
Jon S. Brazier ◽  
Piotr Obuch-Woszczatyński ◽  
Dorota Wultańska ◽  
Felicja Meisel-Mikołajczyk ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
University Hospital ◽  
Erythromycin Resistance ◽  
Toxin B ◽  
Pcr Ribotyping ◽  
Methylase Gene ◽  
High Level ◽  
The University ◽  
Antibiotic Associated Diarrhoea ◽  
Level Resistance

Isolates (79 in total) of Clostridium difficile obtained over a 2 year period from 785 patients suspected of having C. difficile-associated diarrhoea (CDAD) and being hospitalized in the University Hospital in Warsaw were characterized by toxigenicity profile and PCR ribotyping. Furthermore, their susceptibility to clindamycin and erythromycin was determined. Among the 79 C. difficile isolates, 35 were classified as A+B+, 1 as A+B+CDT+, 36 as A−B+ and 7 as A−B−. A total of 21 different PCR ribotypes was detected. Two main A+B+ strains circulated in our hospital: ribotype 014 and ribotype 046. Unexpectedly, the predominant PCR ribotype was type 017, a known A−B+ strain, and this accounted for about 45·5 % of all isolates cultured from patients with CDAD. Isolates belonging to PCR ribotype 017 were found in cases from epidemics of antibiotic-associated diarrhoea in the internal and surgery units. High-level resistance (MIC⩾256 mg l−1) to clindamycin and erythromycin was found in 39 (49 %) of the C. difficile isolates. Interestingly, 34 (94 %) of macrolide-lincosamide-streptogramin B (MLSB) type resistance strains did not produce toxin A, but produced toxin B and were A−B+ ribotype 017. Thirty-seven of the high-level resistance strains harboured the erythromycin-resistance methylase gene (ermB). C. difficile isolates (2/29) that had high-level clindamycin and erythromycin resistance, and belonged to PCR ribotype 046, were ermB negative. These investigations revealed that the predominant C. difficile strain isolated from symptomatic patients hospitalized in University Hospital in Warsaw was MLSB-positive clindamycin/erythromycin-resistant PCR ribotype 017.

scholarly journals A case of Clostridium difficile-associated disease due to the highly virulent clone of Clostridium difficile PCR ribotype 027, March 2007 in Germany

2007 ◽  
Vol 12 (46) ◽  
Author(s):  
N H Zaiss ◽  
J Weile ◽  
G Ackermann ◽  
E J Kuijper ◽  
W Witte ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Pseudomembranous Colitis ◽  
Ribotype 027 ◽  
Pcr Ribotyping ◽  
Pcr Ribotype 027 ◽  
Highly Virulent

Here, we report the isolation of C. difficile PCR ribotype 027 from a patient suffering from pseudomembranous colitis in Germany in March 2007. The strain was identified during a retrospective PCR ribotyping survey of stored isolates.

scholarly journals Advances in molecular surveillance of Clostridium difficile in Bulgaria

2013 ◽  
Vol 62 (9) ◽  
pp. 1428-1434 ◽  
Author(s):  
Elina G. Dobreva ◽  
Ivan N. Ivanov ◽  
Rossitza S. Vathcheva-Dobrevska ◽  
Katucha I. Ivanova ◽  
Galina D. Asseva ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Real Time ◽  
Real Time Pcr ◽  
Simultaneous Detection ◽  
Variable Number ◽  
Innovative Methods ◽  
Pcr Ribotyping ◽  
Typing Methods ◽  
Stool Samples

The increasing incidence of Clostridium difficile infection (CDI) in Bulgaria has indicated the need to implement better surveillance approaches. The aim of the present work was to improve the current surveillance of CDI in Bulgaria by introducing innovative methods for identification and typing. One hundred and twenty stool samples obtained from 108 patients were studied over 4 years from which 32 C. difficile isolates were obtained. An innovative duplex EvaGreen real-time PCR assay based on simultaneous detection of the gluD and tcdB genes was developed for rapid C. difficile identification. Four toxigenic profiles were distinguished by PCR: A+B+CDT− (53.1 %, 17/32), A−B+CDT− (28.1 %, 9/32), A+B+CDT+ (9.4 %, 3/32) and A−B−CDT− (9.4 %, 3/32). PCR ribotyping and multilocus variable number of tandem repeat analysis (MLVA7) were used for molecular characterization of the isolates. In total, nine distinct ribotypes were confirmed and the most prevalent for Bulgarian hospitals was 017 followed by 014/020, together accounting for 44 % of all isolates. Eighteen per cent of the isolates (6/32) did not match any of the 25 reference ribotypes available in this study. Twenty-four MLVA7 genotypes were detected among the clinical C. difficile isolates, distributed as follows: five for 017 ribotype, two for 014/020, 001, 002, 012 and 046 each, and one each for ribotypes 023, 070 and 078. The correlation between the typing methods was significant and allowed the identification of several clonal complexes. These results suggest that most C. difficile cases in the eight Bulgarian hospitals studied were associated with isolates belonging to the outbreak ribotypes 017 and 014/20, which are widely distributed in Europe. The real-time PCR protocol for simultaneous detection of gluD and tcdB proved to be very effective and improved C. difficile identification and confirmation of clinical C. difficile isolates.

scholarly journals Peptidoglycan analysis reveals that synergistic deacetylase activity in vegetative Clostridium difficile impacts the host response

2020 ◽  
Vol 295 (49) ◽  
pp. 16785-16796
Author(s):  
Héloise Coullon ◽  
Aline Rifflet ◽  
Richard Wheeler ◽  
Claire Janoir ◽  
Ivo G. Boneca ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Pathogenic Bacteria ◽  
Pseudomembranous Colitis ◽  
Physiological Role ◽  
Innate Immune ◽  
Innate Immune Responses ◽  
Important Target ◽  
Colonization Ability ◽  
Activity Of Enzymes ◽  
High Level

Clostridium difficile is an anaerobic and spore-forming bacterium responsible for 15–25% of postantibiotic diarrhea and 95% of pseudomembranous colitis. Peptidoglycan is a crucial element of the bacterial cell wall that is exposed to the host, making it an important target for the innate immune system. The C. difficile peptidoglycan is largely N-deacetylated on its glucosamine (93% of muropeptides) through the activity of enzymes known as N-deacetylases, and this N-deacetylation modulates host–pathogen interactions, such as resistance to the bacteriolytic activity of lysozyme, virulence, and host innate immune responses. C. difficile genome analysis showed that 12 genes potentially encode N-deacetylases; however, which of these N-deacetylases are involved in peptidoglycan N-deacetylation remains unknown. Here, we report the enzymes responsible for peptidoglycan N-deacetylation and their respective regulation. Through peptidoglycan analysis of several mutants, we found that the N-deacetylases PdaV and PgdA act in synergy. Together they are responsible for the high level of peptidoglycan N-deacetylation in C. difficile and the consequent resistance to lysozyme. We also characterized a third enzyme, PgdB, as a glucosamine N-deacetylase. However, its impact on N-deacetylation and lysozyme resistance is limited, and its physiological role remains to be dissected. Finally, given the influence of peptidoglycan N-deacetylation on host defense against pathogens, we investigated the virulence and colonization ability of the mutants. Unlike what has been shown in other pathogenic bacteria, a lack of N-deacetylation in C. difficile is not linked to a decrease in virulence.

scholarly journals Intrinsic Class D β-Lactamases of Clostridium difficile

mBio ◽  
2018 ◽  
Vol 9 (6) ◽  
Author(s):  
Marta Toth ◽  
Nichole K. Stewart ◽  
Clyde Smith ◽  
Sergei B. Vakulenko
Keyword(s):  
Clostridium Difficile ◽  
Broad Spectrum ◽  
Antimicrobial Agents ◽  
Catalytic Efficiency ◽  
Content Type ◽  
Mechanism Of Resistance ◽  
Class D ◽  
Infection Resistance ◽  
High Level ◽  
Clostridium Cochlearium

ABSTRACT Clostridium difficile is the causative agent of the deadly C. difficile infection. Resistance of the pathogen to β-lactam antibiotics plays a major role in the development of the disease, but the mechanism of resistance is currently unknown. We discovered that C. difficile encodes class D β-lactamases, i.e., CDDs, which are intrinsic to this species. We studied two CDD enzymes, CDD-1 and CDD-2, and showed that they display broad-spectrum, high catalytic efficiency against various β-lactam antibiotics, including penicillins and expanded-spectrum cephalosporins. We demonstrated that the cdd genes are poorly expressed under the control of their own promoters and contribute only partially to the observed resistance to β-lactams. However, when the cdd1 gene was expressed under the control of efficient promoters in the antibiotic-sensitive Clostridium cochlearium strain, it produced high-level resistance to β-lactams. Taken together, the results determined in this work demonstrate the existence in C. difficile of intrinsic class D β-lactamases which constitute a reservoir of highly potent enzymes capable of conferring broad-spectrum, clinically relevant levels of resistance to β-lactam antibiotics. This discovery is a significant contribution to elucidation of the mechanism(s) of resistance of the clinically important pathogen C. difficile to β-lactam antibiotics. IMPORTANCE C. difficile is a spore-forming anaerobic bacterium which causes infection of the large intestine with high mortality rates. The C. difficile infection is difficult to prevent and treat, as the pathogen is resistant to many antimicrobial agents. Prolonged use of β-lactam antibiotics for treatment of various infectious diseases triggers the infection, as these drugs suppress the abundance of protective gut bacteria, allowing the resistant C. difficile bacteria to multiply. While resistance of C. difficile to β-lactam antibiotics plays the major role in the development of the disease, the mechanism of resistance is unknown. The significance of our research is in the discovery in C. difficile of β-lactamases, enzymes that destroy β-lactam antibiotics. These findings ultimately can help to combat deadly C. difficile infections.

scholarly journals Clostridium difficile infection: a Serbian single-center experience

2015 ◽  
Vol 9 (02) ◽  
pp. 136-140 ◽  
Author(s):  
Milos Korac ◽  
Ivana Milosevic ◽  
Marko Markovic ◽  
Natasa Popovic ◽  
Milena Ilic ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Clostridium Difficile Infection ◽  
Pseudomembranous Colitis ◽  
Antimicrobial Agents ◽  
Stool Culture ◽  
University Hospital ◽  
Watery Diarrhea ◽  
Surgical Interventions ◽  
The Mean ◽  
Hospital Acquired

Introduction: Clostridium difficile infection (CDI) is the most common cause of hospital-acquired diarrhea. Severity of CDI is associated with advanced age and co-morbidities. The clinical spectrum varies from mild watery diarrhea to severe fulminant pseudomembranous colitis with complications. Methodology: This study conducted over a six-year period (2008 to 2013) included 510 patients treated at the University Hospital for Infectious and Tropical Diseases in Belgrade, Serbia. In patients with a history of previous hospitalization and/or treatment with antimicrobial agents who developed diarrhea, the diagnosis was established with rapid tests for C. difficile toxin A and B and by stool culture for C. difficile (454 patients) or by endoscopic examination and histological analyses of the biopsy samples taken from the colonic mucosa (56 patients). Results: The mean age of patients was 67.71± 13.34 years. A total of 67.8% patients were older than 65 years. Over half (58.7%) of the patients were female. 93% had been previously hospitalized and/or had surgical interventions, during which they had been treated with antibiotics. In the clinical presentation spectrum, pseudomembranous colitis occurred in 51.0% .The mean duration of illness after the introduction of specific antibiotic therapy was 7.10 ± 4.88 days. Complications developed in 14 patients. The disease relapsed in 43 (8.4%). Thirty-two (6.3%) patients died, mostly due to co-morbidities. Conclusions: CDI is the most important cause of hospital-acquired diarrhea in Serbia. The disease mainly affects elderly patients with co-morbidities. The incidence of complications is low and prognosis is age dependent and related to pre-existing diseases.

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