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E J Kuijper - One of the best experts on this subject based on the ideXlab platform.
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the emergence of clostridium difficile Ribotypes 027 and 176 with a predominance of the clostridium difficile ribotype 001 recognized in slovakia following the european standardized clostridium difficile infection surveillance of 2016
International Journal of Infectious Diseases, 2020Co-Authors: Elena Novakova, E J Kuijper, Maria Stefkovicova, Maria Garabasova Kopilec, Martin Novak, Nina Kotlebova, Marcela KrutovaAbstract:Abstract Aim To obtain standardized epidemiological data for Clostridium difficile infection (CDI) in Slovakia. Methods Between October and December 2016, 36 hospitals in Slovakia used the European Centre for Disease Prevention and Control (ECDC) Clostridium difficile infection (CDI) surveillance protocol. Results The overall mean CDI incidence density was 2.8 (95% confidence interval 1.9–3.9) cases per 10 000 patient-days. Of 332 CDI cases, 273 (84.9%) were healthcare-associated, 45 (15.1%) were community-associated, and 14 (4.2%) were cases of recurrent CDI. A complicated course of CDI was reported in 14.8% of cases (n = 51). CDI outcome data were available for 95.5% of cases (n = 317). Of the 35 patients (11.1%) who died, 34 did so within 30 days after their CDI diagnosis. Of the 78 isolates obtained from 12 hospitals, 46 belonged to PCR ribotype 001 (59.0%; 11 hospitals) and 23 belonged to ribotype 176 (29.5%; six hospitals). A total of 73 isolates (93.6%) showed reduced susceptibility to moxifloxacin (Ribotypes 001 and 176; p Conclusions These results show the emergence of C. difficile Ribotypes 027 and 176 with a predominance of ribotype 001 in Slovakia in 2016. Given that an almost homogeneous reduced susceptibility to moxifloxacin was detected in C. difficile isolates, this stresses the importance of reducing fluoroquinolone prescriptions in Slovak healthcare settings.
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recreational sandboxes for children and dogs can be a source of epidemic Ribotypes of clostridium difficile
Zoonoses and Public Health, 2018Co-Authors: Cristina Orden, E J Kuijper, Carlos Neila, Jose L Blanco, Sergio Alvarezperez, Celine Harmanus, Marta E GarciaAbstract:Summary Different studies have suggested that the sand of public playgrounds could have a role in the transmission of infections, particularly in children. Furthermore, free access of pets and other animals to the playgrounds might increase such a risk. We studied the presence of Clostridium difficile in 20 pairs of sandboxes for children and dogs located in different playgrounds within the Madrid region (Spain). Clostridium difficile isolation was performed by enrichment and selective culture procedures. The genetic (ribotype and amplified fragment length polymorphism [AFLP]) diversity and antibiotic susceptibility of isolates was also studied. Overall, 52.5% (21/40) of samples were positive for the presence of C. difficile. Eight of the 20 available isolates belonged to the toxigenic Ribotypes 014 (n = 5) and 106 (n = 2), both regarded as epidemic, and CD047 (n = 1). The other 12 isolates were non-toxigenic, and belonged to Ribotypes 009 (n = 5), 039 (n = 4), and 067, 151 and CD048 (one isolate each). Nevertheless, all isolates (even those of a same ribotype) were classified into different AFLP genotypes indicating non-relatedness. In conclusion, our results revealed the presence of epidemic Ribotypes of C. difficile in children's and dog's sandboxes located nearby, which constitutes a major health risk.
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effectiveness of various cleaning and disinfectant products on clostridium difficile spores of pcr Ribotypes 010 014 and 027
Antimicrobial Resistance and Infection Control, 2017Co-Authors: Nikki Kenters, E J Kuijper, Elisabeth G W Huijskens, S C J De Wit, I G J M Sanders, J Van Rosmalen, Andreas VossAbstract:In healthcare facilities, Clostridium difficile infections spread by transmission of bacterial spores. Appropriate sporicidal disinfectants are needed to prevent development of clusters and outbreaks. In this study different cleaning/disinfecting wipes and sprays were tested for their efficacy against spores of distinctive C. difficile PCR Ribotypes. Four different products were tested; 1) hydrogen peroxide 1.5%; 2) glucoprotamin 1.5%; 3) a mixture of ethanol, propane and N-alkyl amino propyl glycine; and 4) a mixture of didecyldimonium chloride, benzalkonium chloride, polyaminopropyl, biguanide and dimenthicone as active ingredients. Tiles were contaminated with a test solution containing a concentration of 5x106CFU/ml spores of C. difficile strains belonging to PCR Ribotypes 010, 014 or 027. The tiles were left to dry for an hour and then wiped or sprayed with one of the sprays or wipes as intended by the manufacturers. When products neutralized after 5 min, microbiological cultures and ATP measures were performed. Irrespective of the disinfection method, the microbial count log10 reduction of C. difficile PCR ribotype 010 was highest, followed by the reduction of C. difficile 014 and C. difficile 027. Overall, the wipes performed better than the sprays with the same active ingredient. On average, although not significantly, a difference in relative light units (RLU) reduction between the wipes and sprays was found. The wipes had a higher RLU log10 reduction, but no significant difference for RLU reduction was observed between the different C. difficile strains (p = 0.16). C. difficile spores of PCR Ribotypes 014 and 027 strains are more difficult to eradicate than non-toxigenic PCR ribotype 010. In general, impregnated cleaning/disinfection wipes performed better than ready-to-use sprays. Wipes with hydrogen peroxide (1.5%) showed the highest bactericidal activity.
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clostridium difficile pcr Ribotypes 001 and 176 the common denominator of c difficile infection epidemiology in the czech republic 2014
Eurosurveillance, 2016Co-Authors: Marcela Krutova, E J Kuijper, Jana Matejkova, Pavel Drevinek, Otakar NycAbstract:In 2014, 18 hospitals in the Czech Republic participated in a survey of the incidence of Clostridium difficile infections (CDI) in the country. The mean CDI incidence was 6.1 (standard deviation (SD):7.2) cases per 10,000 patient bed-days and 37.8 cases (SD: 41.4) per 10,000 admissions. The mean CDI testing frequency was 39.5 tests (SD: 25.4) per 10,000 patient bed-days and 255.8 tests (SD: 164.0) per 10,000 admissions. A total of 774 C. difficile isolates were investigated, of which 225 (29%) belonged to PCR ribotype 176, and 184 isolates (24%) belonged to PCR ribotype 001. Multilocus variable-number tandem repeat analysis (MLVA) revealed 27 clonal complexes formed by 84% (190/225) of PCR ribotype 176 isolates, and 14 clonal complexes formed by 77% (141/184) of PCR ribotype 001 isolates. Clonal clusters of PCR Ribotypes 176 and 001 were observed in 11 and 7 hospitals, respectively. Our data demonstrate the spread of two C. difficile PCR Ribotypes within 18 hospitals in the Czech Republic, stressing the importance of standardising CDI testing protocols and implementing mandatory CDI surveillance in the country.
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pcr ribotype prevalence and molecular basis of macrolide lincosamide streptogramin b mlsb and fluoroquinolone resistance in irish clinical clostridium difficile isolates
Journal of Antimicrobial Chemotherapy, 2011Co-Authors: K Solomon, Seamus Fanning, Sinead Mcdermott, Sean Murray, Louise Scott, Alan Martin, Mairead Skally, Karen Burns, E J KuijperAbstract:Background Antimicrobial use is recognized as a risk factor for Clostridium difficile infection (CDI) and outbreaks. We studied the relationship between PCR ribotype, antimicrobial susceptibility and the genetic basis of resistance in response to exposure to antimicrobial agents. Methods C. difficile isolates were cultured from 133 CDI patients for whom recent antimicrobial drug exposure had been recorded. Isolates were ribotyped by PCR and assessed for their susceptibility to the macrolide-lincosamide-streptogramin B (MLS(B)) group of compounds (erythromycin and clindamycin) and fluoroquinolone antimicrobials (ciprofloxacin, levofloxacin and moxifloxacin). Where relevant, the genetic basis of resistance was determined. Results Prevalent Ribotypes (including 027, 001 and 106) exhibited significantly greater antimicrobial resistance compared with Ribotypes 078 and 014, among others. Clindamycin-resistant ribotype 078 was detected for the first time. Ribotypes 027 and 001 were more likely to exhibit MLS(B) resistance, a feature that was associated with the erm(B) gene. Exposure to MLS(B) or fluoroquinolone antimicrobial compounds in the 8 weeks prior to the onset of infection was not associated with specific genetic markers of resistance. Single amino acid substitutions in the A and B subunits of DNA gyrase were noted and were ribotype specific and linked to resistance to moxifloxacin. Conclusions Resistance to MLS(B) and fluoroquinolone antimicrobial compounds is common among prevalent Ribotypes of C. difficile. The genetic basis for antimicrobial resistance appears to be ribotype specific and conserved in the absence of recent antimicrobial selection pressure.
Mark H Wilcox - One of the best experts on this subject based on the ideXlab platform.
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diversity of clostridium difficile pcr Ribotypes in europe results from the european multicentre prospective biannual point prevalence study of clostridium difficile infection in hospitalised patients with diarrhoea euclid 2012 and 2013
Eurosurveillance, 2016Co-Authors: Kerrie Davies, Helen Ashwin, Christopher Longshaw, David A Burns, Georgina Davis, Mark H WilcoxAbstract:Clostridium difficile infection (CDI) is the major cause of infective diarrhoea in healthcare environments. As part of the EUCLID study, the largest C. difficile epidemiological study of its type, PCR-ribotype distribution of C. difficile isolates in Europe was investigated. PCR-ribotyping was performed on 1196 C. difficile isolates from diarrhoeal samples sent to the European co-ordinating laboratory in 2012–13 by 482 participating hospitals from 19 European countries. A total of 125 Ribotypes were identified, of which Ribotypes 027 (18.6%), 001/072 (11.2%) and 014 (7.0%) were the most prevalent. Distinct regional patterns of ribotype distribution were noted in Northern, Western, Southern and Eastern Europe. Of 596 isolates from patients with toxin-positive stools (confirmed CDI), ribotype 027 accounted for 27.8% of infections in patients aged 2–<65 years, but the prevalence decreased in those aged ≥65 years (14.3%) and further decreased in those aged ≥81 years (9.2%). The prevalence of ribotype 027 (but not other epidemic strains) was inversely proportional to overall ribotype diversity (R2=0.717). The EUCLID study highlights an increased diversity of C. difficile Ribotypes across Europe compared with previous studies, with considerable inter-country variation in ribotype distribution. Continuous surveillance programmes are necessary to monitor the changing epidemiology of C. difficile
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diversity of clostridium difficile pcr Ribotypes in europe results from the european multicentre prospective biannual point prevalence study of clostridium difficile infection in hospitalised patients with diarrhoea euclid 2012 and 2013
Eurosurveillance, 2016Co-Authors: Kerrie Davies, Helen Ashwin, Christopher Longshaw, David A Burns, Georgina Davis, Mark H WilcoxAbstract:: Clostridium difficile infection (CDI) is the major cause of infective diarrhoea in healthcare environments. As part of the European, multicentre, prospective, biannual, point-prevalence study of Clostridium difficile infection in hospitalised patients with diarrhoea (EUCLID), the largest C. difficile epidemiological study of its type, PCR ribotype distribution of C. difficile isolates in Europe was investigated. PCR ribotyping was performed on 1,196 C. difficile isolates from diarrhoeal samples sent to the European coordinating laboratory in 2012-13 and 2013 (from two sampling days) by 482 participating hospitals from 19 European countries. A total of 125 Ribotypes were identified, of which Ribotypes 027 (19%, n =222), 001/072 (11%, n = 134) and 014/020 (10%, n = 119) were the most prevalent. Distinct regional patterns of ribotype distribution were noted. Of 596 isolates from patients with toxin-positive stools (CDI cases), ribotype 027 accounted for 22% (32/144) of infections in cases aged from 18 to less than 65 years, but the prevalence decreased in those aged ≥ 65 years (14% (59/412)) and further decreased in those aged ≥ 81 years (9% (18/195)). The prevalence of ribotype 027 and 176, but not other epidemic strains, was inversely proportional to overall ribotype diversity (R(2) = 0.717). This study highlights an increased diversity of C. difficile Ribotypes across Europe compared with previous studies, with considerable intercountry variation in ribotype distribution. Continuous surveillance programmes are necessary to monitor the changing epidemiology of C. difficile.
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pan european longitudinal surveillance of antibiotic resistance among prevalent clostridium difficile Ribotypes
Clinical Microbiology and Infection, 2015Co-Authors: Jane Freeman, Jonathan Vernon, Kirsti Morris, Scott Nicholson, Sharie L Todhunter, Christopher Longshaw, Mark H WilcoxAbstract:Clostridium difficile infection remains a major healthcare burden. Until the recent introduction of fidaxomicin, antimicrobial treatments were limited to metronidazole and vancomycin. The emergence of epidemic C. difficile PCR ribotype 027 and its potential link to decreased antibiotic susceptibility highlight the lack of large-scale antimicrobial susceptibility and epidemiological data available. We report results of epidemiological and antimicrobial susceptibility investigations of C. difficile isolates collected prior to fidaxomicin introduction, establishing important baseline data. Thirty-nine sites in 22 countries submitted a total of 953 C. difficile isolates for PCR ribotyping, toxin testing, and susceptibility testing to metronidazole, vancomycin, fidaxomicin, rifampicin, moxifloxacin, clindamycin, imipenem, chloramphenicol, and tigecycline. Ninety-nine known Ribotypes were identified. Ribotypes 027, 014, 001/072, and 078 were most frequently isolated in line with previous European studies. There was no evidence of resistance to fidaxomicin, and reduced susceptibility to metronidazole and vancomycin was also scarce. Rifampicin, moxifloxacin, and clindamycin resistance (13%, 40%, and 50% of total isolates, respectively) were evident in multiple Ribotypes. There was a significant correlation between lack of ribotype diversity and greater antimicrobial resistance (measured by cumulative resistance score). Well-known epidemic Ribotypes 027 and 001/072 were associated with multiple antimicrobial resistance, but high levels of resistance were also observed, particularly in 018 and closely related emergent ribotype 356 in Italy. This raises the possibility of antimicrobial exposure as the underlying reason for their appearance, and highlights the need for ongoing epidemiological and antimicrobial resistance surveillance.
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changing epidemiology of clostridium difficile infection following the introduction of a national ribotyping based surveillance scheme in england
Clinical Infectious Diseases, 2012Co-Authors: Mark H Wilcox, Warren N. Fawley, N P Shetty, M Shemko, Pietro G Coen, A Birtles, M Cairns, M D Curran, K J Dodgson, S M GreenAbstract:BACKGROUND: Marked increases in Clostridium difficile infection (CDI) incidence, driven by epidemic strain spread, is a global phenomenon. METHODS: The Clostridium difficile Ribotyping Network (CDRN) was established in 2007 as part of enhanced CDI surveillance in England, to facilitate the recognition and control of epidemic strains. We report on changes in CDI epidemiology in England in the first 3 years of CDRN. RESULTS: CDRN received 12,603 fecal specimens, comprising significantly (P < .05) increasing numbers and proportions of national CDI cases in 2007-2008 (n = 2109, 3.8%), 2008-2009 (n = 4774, 13.2%), and 2009-2010 (n = 5720, 22.3%). The C. difficile recovery rate was 90%, yielding 11,294 isolates for ribotyping. Rates of 9 of the 10 most common Ribotypes changed significantly (P < .05) during 2007-2010. Clostridium difficile ribotype 027 predominated, but decreased markedly from 55% to 36% and 21% in 2007-2008, 2008-2009, and 2009-2010, respectively. The largest regional variations in prevalence occurred for Ribotypes 027, 002, 015, and 078. Cephalosporin and fluoroquinolone use in CDI cases was reported significantly (P < .05) less frequently during 2007-2010. Mortality data were subject to potential reporting bias, but there was a significant decrease in CDI-associated deaths during 2007-2010, which may have been due to multiple factors, including reduced prevalence of ribotype 027. CONCLUSIONS: Access to C. difficile ribotyping was associated with significant changes in the prevalence of epidemic strains, especially ribotype 027. These changes coincided with markedly reduced CDI incidence and related mortality in England. CDI control programs should include prospective access to C. difficile typing and analysis of risk factors for CDI and outcomes.
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emergence of reduced susceptibility to metronidazole in clostridium difficile
Journal of Antimicrobial Chemotherapy, 2008Co-Authors: Simon D Baines, Warren N. Fawley, E J Kuijper, Paola Mastrantonio, Jane Freeman, Celine Harmanus, Rachael Oconnor, Mark H WilcoxAbstract:OBJECTIVES: Antimicrobial treatment for Clostridium difficile infection (CDI) has typically been metronidazole, although reports have questioned the efficacy of this option. We screened recently isolated C. difficile (2005-06) for susceptibility to metronidazole and compared results for historic isolates (1995-2001). METHODS: C. difficile Ribotypes 001 (n = 86), 106 (n = 81) and 027 (n = 48) and isolates from the 10 other most prevalent Ribotypes in Leeds (n = 57) were screened using spiral gradient endpoint analysis (SGE). C. difficile with metronidazole SGE MICs > or = 6 mg/L were analysed further by agar incorporation and Etest. Multiple-locus variable-number tandem-repeat analysis (MLVA) typing was performed for 28 C. difficile isolates. RESULTS: No reduced metronidazole susceptibility was observed in C. difficile Ribotypes 106 and 027 (geometric mean SGE MICs 1.11 and 0.90 mg/L, respectively). In contrast, 21 (24.4%) C. difficile ribotype 001 demonstrated reduced susceptibility to metronidazole (geometric mean SGE MICs 3.51 mg/L, P < 0.001). Variations in susceptibility were observed relating to the method and media, but increased metronidazole MICs were confirmed by an agar incorporation method. Geometric mean agar incorporation MICs for historic C. difficile ribotype 001 (n = 72) were 1.03 (range 0.25-2) mg/L compared with 5.94 (4-8) mg/L (P < 0.001) for recent isolates displaying reduced metronidazole susceptibility. MLVA typing revealed two clonal complexes of C. difficile with reduced susceptibility to metronidazole. CONCLUSIONS: We have demonstrated the emergence of reduced susceptibility to metronidazole in 24.4% of the recent C. difficile ribotype 001 isolates from our institution. Our observations could have implications in the clinical setting due to the poor penetration of metronidazole into the colon.
Maja Rupnik - One of the best experts on this subject based on the ideXlab platform.
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distribution of clostridium difficile pcr Ribotypes and high proportion of 027 and 176 in some hospitals in four south eastern european countries
Anaerobe, 2016Co-Authors: Maja Rupnik, Arjana Tambic Andrasevic, Elena Trajkovska Dokic, Ivanka Matas, Milica Jovanovic, Selma Pasic, Aleksander Kocuvan, Sandra JanezicAbstract:While Clostridium difficile epidemiology is well documented in many European countries, data are largely missing for South Eastern European region. Here we report the PCR ribotype distribution of 249 C. difficile isolates received for typing from six hospital settings from Croatia, Bosnia and Herzegovina, Republic of Macedonia and Serbia in time period from 2008 to 2015. Twenty-four PCR Ribotypes were detected. The majority of strains from Bosnia and Herzegovina and Serbia belonged to PCR ribotype 027 (65.8%). Other three dominating PCR Ribotypes were 176 (18 strains; Croatia), 001/072 (15 strains; all countries) and 014/020 (15 strains; all countries).
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highly divergent clostridium difficile strains isolated from the environment
PLOS ONE, 2016Co-Authors: Sandra Janezic, Valerija Zidaric, Mojca Potocnik, Maja RupnikAbstract:Clostridium difficile is one of the most important human and animal pathogens. However, the bacterium is ubiquitous and can be isolated from various sources. Here we report the prevalence and characterization of C. difficile in less studied environmental samples, puddle water (n = 104) and soil (n = 79). C. difficile was detected in 14.4% of puddle water and in 36.7% of soil samples. Environmental strains displayed antimicrobial resistance patterns comparable to already published data of human and animal isolates. A total of 480 isolates were grouped into 34 different PCR Ribotypes. More than half of these (52.9%; 18 of 34) were already described in humans or animals. However, 14 PCR Ribotypes were new in our PCR ribotype library and all but one were non-toxigenic. The multilocus sequence analysis of these new PCR Ribotypes revealed that non-toxigenic environmental isolates are phylogenetically distinct and belong to three highly divergent clades, two of which have not been described before. Our data suggest that environment is a potential reservoir of genetically diverse population of C. difficile.
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Sporulation properties and antimicrobial susceptibility in endemic and rare Clostridium difficile PCR Ribotypes
Anaerobe, 2016Co-Authors: Valerija Zidaric, Maja RupnikAbstract:Increased sporulation and antibiotic resistance have been proposed to be associated with certain Clostridium difficile epidemic strains such as PCR ribotype 027. In this study we examined these properties in another widespread PCR ribotype, 014/020, in comparison to prevalent PCR ribotype 002 and a group of rarely represented PCR Ribotypes. Highest sporulation was observed in 014/020 strains at 24 h, while after 72 h PCR ribotype 002 and rare PCR Ribotypes formed higher total number of spores. PCR ribotype 014/020 strains exhibited slightly higher resistance to tested antimicrobials, followed by group of rare PCR Ribotypes and less common PCR ribotype 002. Neither sporulation properties nor antibiotic resistance clearly differed in endemic and rare strains.
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Clostridium difficile Ribotypes in humans and animals in Brazil
Memórias do Instituto Oswaldo Cruz, 2015Co-Authors: Rodrigo Otávio Silveira Silva, Maja Rupnik, Amanda Nádia Diniz, Eduardo Garcia Vilela, Francisco Carlos Faria LobatoAbstract:Clostridium difficile is an emerging enteropathogen responsible for pseudomembranous colitis in humans and diarrhoea in several domestic and wild animal species. Despite its known importance, there are few studies aboutC. difficile polymerase chain reaction (PCR) Ribotypes in Brazil and the actual knowledge is restricted to studies on human isolates. The aim of the study was therefore to compare C. difficileRibotypes isolated from humans and animals in Brazil. Seventy-six C. difficile strains isolated from humans (n = 25), dogs (n = 23), piglets (n = 12), foals (n = 7), calves (n = 7), one cat, and one manned wolf were distributed into 24 different PCR Ribotypes. Among toxigenic strains, PCR Ribotypes 014/020 and 106 were the most common, accounting for 14 (18.4%) and eight (10.5%) samples, respectively. Fourteen different PCR Ribotypes were detected among human isolates, nine of them have also been identified in at least one animal species. PCR ribotype 027 was not detected, whereas 078 were found only in foals. This data suggests a high diversity of PCR Ribotypes in humans and animals in Brazil and support the discussion of C. difficile as a zoonotic pathogen.
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Carriage of Clostridium difficile in free-living South American coati (Nasua nasua) in Brazil
Anaerobe, 2014Co-Authors: Rodrigo Otávio Silveira Silva, Lara Ribeiro De Almeida, Carlos Augusto Oliveira Junior, Danielle Ferreira De Magalhães Soares, Pedro Lúcio Lithg Pereira, Maja Rupnik, Francisco Carlos Faria LobatoAbstract:Abstract The objective of this study was to isolate and characterize Clostridium difficile strains in stool samples from a wild urban mammal, a South American coati (Nasua nasua) in Brazil. Forty-six free-living N. nasua were trapped, and stool samples were collected. C. difficile was isolated from three (6.5%) sampled animals, two strains were toxigenic (A+B+CDT−, PCR ribotype 014/020 and 106) and one was non toxigenic (A−B−CDT−, PCR ribotype 053). The present work confirms that ring-tailed coati (N. nasua) could harbor C. difficile strains, including those PCR Ribotypes commonly reported in C. difficile infection in humans.
Gary French - One of the best experts on this subject based on the ideXlab platform.
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lack of effect of strain type on detection of toxigenic clostridium difficile by glutamate dehydrogenase and polymerase chain reaction
Diagnostic Microbiology and Infectious Disease, 2011Co-Authors: Simon D Goldenberg, Maria Gumban, Anthony Hall, Amita Patel, Gary FrenchAbstract:Glutamate dehydrogenase (GDH) is popular as a preliminary test for the detection of Clostridium difficile. Recent work has suggested that GDH sensitivity may vary according to ribotype and may be lower for Ribotypes 002, 027, and 106 compared with polymerase chain reaction (PCR). We investigated this effect using a dilution series of 64 isolates tested by GDH and Cepheid GeneXpert PCR. PCR was significantly more sensitive than GDH overall; however, there was no difference in detection according to specific ribotype.
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lack of association of tcdc type and binary toxin status with disease severity and outcome in toxigenic clostridium difficile
Journal of Infection, 2011Co-Authors: Simon D Goldenberg, Gary FrenchAbstract:Summary The production of binary toxin and presence of truncating mutations in the putative toxin repressor gene, tcdC , have been associated with the increased virulence and spread of Clostridium difficile, especially ribotype 027. We analysed the prevalence of binary toxin genes and tcdC mutations in 207 clinical C. difficile isolates collected between 2008–2010. The majority (83%) belonged to one of five tcdC types and 8% were ribotype 027. There was little evidence of epidemic spread but there was a high prevalence of both predicted tcdC truncating mutations (15%) and binary toxin genes (28%), which occurred in both 027 and other Ribotypes. We measured risk factors (age and laboratory markers) and patient outcomes (severity of disease, ICU admission, mortality, recurrence and length of stay) for patients infected with C. difficile strains with and without these mutations and genes. There was a significantly higher serum C-reactive protein and total peripheral white cell count in the group with predicted tcdC truncating mutations, but no difference in patient outcome. The group with binary toxin genes had a significantly higher total peripheral white cell count and 30 day all cause mortality. We have demonstrated a high prevalence of both predicted tcdC truncating mutations and binary toxin genes in a variety of C. difficile Ribotypes, however neither of these factors by themselves predicted clinical virulence. This and other work show that commonly described deletions and truncating mutations do not by themselves explain the virulence of ribotype 027 and other C. difficile strains and further work is required to explain why some isolates appear to produce more severe disease than others.
J S Brazier - One of the best experts on this subject based on the ideXlab platform.
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extended multilocus variable number tandem repeat analysis of clostridium difficile correlates exactly with ribotyping and enables identification of hospital transmission
Journal of Clinical Microbiology, 2011Co-Authors: Susan Manzoor, Peter M Hawkey, Hannah E Tanner, Clare Marriott, J S Brazier, Katie Hardy, S PlattAbstract:PCR ribotyping is currently used in many countries for epidemiological investigation to track transmission and to identify emerging variants of Clostridium difficile. Although PCR ribotyping differentiates over 300 types, it is not always sufficiently discriminatory for epidemiological investigations particularly for common Ribotypes, e.g., Ribotypes 027, 106, and 017. Multilocus variable-number tandem-repeat analysis (MLVA) is a highly discriminatory molecular subtyping method that has been applied to a number of bacterial species for high-level subtyping. Two MLVA typing schemes for C. difficile have been previously published, each utilizing seven variable-number tandem-repeat (VNTR) loci on the genome with four loci common to both schemes. Although these schemes are good genotyping methods with the ability to discriminate between isolates, they do not identify the ribotype. We show here that increasing the number of VNTR loci to 15, creating the extended MLVA (eMLVA) scheme, we have successfully subtyped all clinically significant Ribotypes while still clustering isolates in concordance with PCR ribotyping. The eMLVA scheme developed here provides insight into the genetic diversity of the C. difficile population at both global and cross-infection clusters in patient levels, with the possibility of replacing PCR ribotyping.
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distribution and antimicrobial susceptibility patterns of clostridium difficile pcr Ribotypes in english hospitals 2007 08
Eurosurveillance, 2008Co-Authors: J S Brazier, R Raybould, Bharat Patel, G Duckworth, A Pearson, Andre Charlett, B I DuerdenAbstract:A surveillance study designed to provide a representative sample of the strains of Clostridium difficile causing infections in hospitals in England was in operation from April 2007 to the end of March 2008. Six hundred and seventy-seven isolates were obtained from 186 hospitals in the nine geographical regions of England as recognised by the Health Protection Agency's Regional Microbiology Network. Typing studies revealed that PCR ribotype 027 is now the most common strain isolated from symptomatic patients, accounting for over 41.3% of isolates in English hospitals. Type 106 was the second most common strain (20.2%) and Type 001, which was once the most common strain associated with hospital outbreaks, has now been reduced to only 7.8% of the total. A mixture of 44 other PCR Ribotypes accounted for the remaining 28.9% of isolates. This represents a changing distribution of strains when compared to a previous study performed two years earlier which showed roughly equal proportions of types 106, 001 and 027. Antimicrobial susceptibility testing by the E test method revealed significantly lower susceptibility to metronidazole in the more common strains when compared to the less common Ribotypes, although none were classified as clinically resistant. Similarly, no resistance to vancomycin was detected. However, common PCR Ribotypes were more resistant to moxifloxacin and erythromycin than the less common strains, which may indicate a selective advantage for resistance to these agents, and combined resistance to these two agents was a good indicator of a common ribotype.
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prevalence of pcr Ribotypes among clostridium difficile isolates from pigs calves and other species
Journal of Clinical Microbiology, 2007Co-Authors: Kevin M Keel, J S Brazier, Karen W Post, Scott Weese, Glenn J SongerAbstract:PCR Ribotypes were obtained for 144 Clostridium difficile isolates from neonatal pigs. Porcine isolates comprised four PCR Ribotypes, but one, ribotype 078, predominated (83%). This was also the most common ribotype (94%) among 33 calf isolates but was rarely identified in other species.
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prevalence and association of pcr Ribotypes of clostridium difficile isolated from symptomatic patients from warsaw with macrolide lincosamide streptogramin b mlsb type resistance
Journal of Medical Microbiology, 2006Co-Authors: Hanna Pituch, J S Brazier, Piotr Obuchwoszczatynski, Dorota Wultanska, F Meiselmikolajczyk, Miroslaw łuczakAbstract: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.
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prevalent pcr Ribotypes of clinical and environmental strains of clostridium difficile isolated from intensive therapy unit patients in kuwait
Journal of Medical Microbiology, 2003Co-Authors: V O Rotimi, J S Brazier, Wafaa Jamal, Eiman M Mokaddas, Molly Johny, B I DuerdenAbstract:Ninety-five isolates of Clostridium difficile from symptomatic and asymptomatic patients and 18 from their environment in the intensive-therapy units (ITUs) of four teaching hospitals in Kuwait were typed by PCR amplification of rRNA intergenic spacer regions (PCR ribotyping). A total of 32 different Ribotypes was detected among the clinical isolates. The predominant Ribotypes from the clinical isolates were types 097 and 078, which accounted for ∼ 40 % of all isolates in the ITUs in Kuwait. Ribotypes 097 (toxigenic), 078 (toxigenic) and 039 (non-toxigenic) were three distinct clones that were circulating in all four hospitals. Ribotypes 097, 078 and 076 (i.e. 50 % of isolates from symptomatic patients) were the predominant isolates associated with C. difficile-associated disease (CDAD). The environmental isolates belonged to a diverse range of Ribotypes, with no particular types common to all the hospitals. Ribotype 078 was found only in the patient environment in Mubarak hospital, while ribotype 097 was restricted to Amiri hospital. The hospital environment occupied by symptomatic as well as symptom-free patients was contaminated with C. difficile. Eight new strains that did not match any in the PCR ribotype library established at the PHLS Anaerobe Reference Unit, Cardiff, UK, were assigned Ribotypes 105, 125, 128, 129, 131, 134, 140 and 141. These findings show that the isolates associated with CDAD in Kuwait are different from those found in the UK and some other European countries.
