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 Novel Complex Mutant β-Lactamase, TEM-68, Identified in a Klebsiella pneumoniae Isolate from an Outbreak of Extended-Spectrum β-Lactamase-Producing Klebsiellae

2000 ◽  
Vol 44 (6) ◽  
pp. 1499-1505 ◽  
Author(s):  
Janusz Fiett ◽  
Andrzej Pałucha ◽  
Beata Mia˛czyńska ◽  
Maria Stankiewicz ◽  
Hanna Przondo-Mordarska ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
University Hospital ◽  
Pediatric Hospitals ◽  
Extended Spectrum ◽  
Novel Complex ◽  
High Level ◽  
Inhibitor Resistance ◽  
The University ◽  
Level Resistance

ABSTRACT Twenty-two Klebsiella pneumoniae and two K. oxytoca extended-spectrum β-lactamase (ESBL)-producing isolates were collected in 1996 from patients in two pediatric wards of the University Hospital in Wrocław, Poland. Molecular typing has revealed that the K. pneumoniae isolates represented four different epidemic strains. Three kinds of enzymes with ESBL activity (pI values of 5.7, 6.0, and 8.2) were identified. The pI 6.0 β-lactamases belonged to the TEM family, and sequencing of thebla TEM genes amplified from representative isolates revealed that these enzymes were TEM-47, previously identified in K. pneumoniae isolates from pediatric hospitals in Łódź and Warsaw. One of the TEM-47-producing strains from Wrocław was very closely related to the isolates from the other cities, and this indicated countrywide spread of the epidemic strain. The pI 5.7 β-lactamase was produced by a single K. pneumoniae isolate for which, apart from oxyimino-β-lactams, the MICs of β-lactam–inhibitor combinations were also remarkably high. Sequencing revealed that this was a novel TEM β-lactamase variant, TEM-68, specified by the following combination of mutations: Gly238Ser, Glu240Lys, Thr265Met, and Arg275Leu. The new enzyme has most probably evolved from TEM-47 by acquiring the single substitution of Arg275, which before was identified only twice in enzymes with inhibitor resistance (IR) activity. TEM-68 was shown to be a novel complex mutant TEM β-lactamase (CMT-2) which combines strong ESBL activity with relatively weak IR activity and, when expressed inK. pneumoniae, is able to confer high-level resistance to a wide variety of β-lactams, including inhibitor combinations. This data confirms the role of the Arg275Leu mutation in determining IR activity and documents the first isolation of K. pneumoniae producing the complex mutant enzyme.

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.

scholarly journals Epidemic potential and antimicrobial resistance in Clostridium difficile

2007 ◽  
Vol 28 (4) ◽  
pp. 195
Author(s):  
Thomas V Riley
Keyword(s):  
Clostridium Difficile ◽  
Financial Burden ◽  
Hospital Environment ◽  
Comorbid Conditions ◽  
Subsequent Infection ◽  
Toxin B ◽  
Exposure Age ◽  
Hospital Acquired ◽  
Hospital Inpatients ◽  
Antibiotic Associated Diarrhoea

Clostridium difficile is the most commonly diagnosed cause of infectious hospital-acquired diarrhoea. C. difficile was first isolated in 1935 but not identified as the main causative agent of antibiotic-associated diarrhoea (AAD) and pseudomemranous colitis (PMC) until 1977. The spectrum of disease caused by C. difficile ranges from asymptomatic colonisation to colitis that can progress to more severe PMC. Complications include colonic perforation and death. The term C. difficile-associated diarrhoea (CDAD) is used to describe the symptomatic manifestations of the disease, thus excluding asymptomatic colonisation. Hospital inpatients with CDAD are generally elderly and have several comorbid conditions. The majority of these patients have been exposed to antimicrobials that reduce ?colonisation resistance? of the large intestine allowing subsequent infection with C. difficile. Whether infection progresses to disease is determined by many factors such as antibiotic exposure, age and comorbidities, and others that are as yet unknown. Acquisition of C. difficile is facilitated by its ability to form spores that are resistant to many disinfectants, allowing it to remain viable in the hospital environment for long periods of time. Toxigenic isolates of C. difficile usually produce two toxins, toxin A and toxin B, and these are thought of as the major virulence factors. CDAD is a major financial burden on healthcare systems, with patients spending an extra one?three weeks in hospital costing US$5?10,000 per episode.

scholarly journals Phenotypes and Genotypes of Erythromycin-ResistantStreptococcus pyogenes Strains in Italy and Heterogeneity of Inducibly Resistant Strains

1999 ◽  
Vol 43 (8) ◽  
pp. 1935-1940 ◽  
Author(s):  
Eleonora Giovanetti ◽  
Maria Pia Montanari ◽  
Marina Mingoia ◽  
Pietro Emanuele Varaldo
Keyword(s):  
Streptococcus Pyogenes ◽  
Tetracycline Resistance ◽  
Erythromycin Resistance ◽  
Testing Conditions ◽  
Resistant Strains ◽  
Methylase Gene ◽  
Set Up ◽  
Susceptibility Patterns ◽  
Disk Test ◽  
Level Resistance

ABSTRACT A total of 387 clinical strains of erythromycin-resistant (MIC, ≥1 μg/ml) Streptococcus pyogenes, all isolated in Italian laboratories from 1995 to 1998, were examined. By the erythromycin-clindamycin double-disk test, 203 (52.5%) strains were assigned to the recently described M phenotype, 120 (31.0%) were assigned to the inducible macrolide, lincosamide, and streptogramin B resistance (iMLS) phenotype, and 64 (16.5%) were assigned to the constitutive MLS resistance (cMLS) phenotype. The inducible character of the resistance of the iMLS strains was confirmed by comparing the clindamycin MICs determined under normal testing conditions and those determined after induction by pregrowth in 0.05 μg of erythromycin per ml. The MICs of erythromycin, clarithromycin, azithromycin, josamycin, spiramycin, and the ketolide HMR3004 were then determined and compared. Homogeneous susceptibility patterns were observed for the isolates of the cMLS phenotype (for all but one of the strains, HMR3004 MICs were 0.5 to 8 μg/ml and the MICs of the other drugs were >128 μg/ml) and those of the M phenotype (resistance only to the 14- and 15-membered macrolides was recorded, with MICs of 2 to 32 μg/ml). Conversely, heterogeneous susceptibility patterns were observed in the isolates of the iMLS phenotype, which were subdivided into three distinct subtypes designated iMLS-A, iMLS-B, and iMLS-C. The iMLS-A strains (n = 84) were highly resistant to the 14-, 15-, and 16-membered macrolides and demonstrated reduced susceptibility to low-level resistance to HMR3004. The iMLS-B strains (n = 12) were highly resistant to the 14- and 15-membered macrolides, susceptible to the 16-membered macrolides (but highly resistant to josamycin after induction), and susceptible to HMR3004 (but intermediate or resistant after induction). The iMLS-C strains (n = 24) had lower levels of resistance to the 14- and 15-membered macrolides (with erythromycin MICs increasing two to four times after induction), were susceptible to the 16-membered macrolides (but resistant to josamycin after induction), and remained susceptible to HMR3004, also after induction. The erythromycin resistance genes in 100 isolates of the different groups were investigated by PCR. All cMLS and iMLS-A isolates tested had theermAM (ermB) gene, whereas all iMLS-B and iMLS-C isolates had the ermTR gene (neither methylase gene was found in isolates of other groups). The M isolates had only the macrolide efflux (mefA) gene, which was also found in variable proportions of cMLS, iMLS-A, iMLS-B, and iMLS-C isolates. The three iMLS subtypes were easily differentiated by a triple-disk test set up by adding a josamycin disk to the erythromycin and clindamycin disks of the conventional double-disk test. Tetracycline resistance was not detected in any isolate of the iMLS-A subtype, whereas it was observed in over 90% of both iMLS-B and iMLS-C isolates.

scholarly journals Fluoroquinolone and Macrolide Exposure Predict Clostridium difficile Infection with the Highly Fluoroquinolone- and Macrolide-Resistant Epidemic C. difficile Strain BI/NAP1/027

2015 ◽  
Vol 60 (1) ◽  
pp. 418-423 ◽  
Author(s):  
Jeffrey T. Wieczorkiewicz ◽  
Bert K. Lopansri ◽  
Adam Cheknis ◽  
James R. Osmolski ◽  
David W. Hecht ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Restriction Endonuclease Analysis ◽  
First Episode ◽  
Retrospective Case ◽  
Content Type ◽  
High Level ◽  
Control Study ◽  
Level Resistance ◽  
Endonuclease Analysis ◽  
Multiple Antibiotics

ABSTRACTAntibiotics have been shown to influence the risk of infection with specificClostridium difficilestrains as well as the risk ofC. difficileinfection (CDI). We performed a retrospective case-control study of patients infected with the epidemic BI/NAP1/027 strain in a U.S. hospital following recognition of increased CDI severity and culture of stools positive byC. difficiletoxin immunoassay. Between 2005 and 2007, 72% (103/143) of patients with first-episode CDIs were infected with the BI strain by restriction endonuclease analysis (REA) typing. Most patients received multiple antibiotics within 6 weeks of CDI onset (median of 3 antibiotic classes). By multivariate analysis, fluoroquinolone and macrolide exposure was more frequent among BI cases than among non-BI-infected controls (odds ratio [OR] for fluoroquinolones, 3.2; 95% confidence interval [CI], 1.3 to 7.5; (P< 0.001; OR for macrolides, 5.2; 95% CI, 1.1 to 24.0;P= 0.04)). In contrast, clindamycin use was less frequent among the BI cases than among the controls (OR, 0.1; 95% CI, 0.03 to 0.4;P= 0.001). High-level resistance to moxifloxacin and azithromycin was more frequent among BI strains (moxifloxacin, 49/102 [48%] BI versus 0/40 non-BI,P= 0.0001; azithromycin, 100/102 [98%] BI versus 22/40 [55%] non-BI,P= 0.0001). High-level resistance to clindamycin was more frequent among non-BI strains (22/40 [55%] non-BI versus 7/102 [7%] BI,P= 0.0001). Fluoroquinolone use, macrolide use, andC. difficileresistance to these antibiotic classes were associated with infection by the epidemic BI strain ofC. difficilein a U.S. hospital during a time when CDI rates were increasing nationally due to the highly fluoroquinolone-resistant BI/NAP1/027 strain.

Outbreak of Infections in a Greek University Hospital Involving a Single Clone of High-Level Aminoglycoside-ResistantEnterococcus faecalis

2000 ◽  
Vol 21 (12) ◽  
pp. 786-789 ◽  
Author(s):  
Spyros Pournaras ◽  
Athanassios Tsakris ◽  
Mary E. Kaufmann ◽  
John Douboyas ◽  
Antonios Antoniadis
Keyword(s):  
Enterococcus Faecalis ◽  
Resistance Genes ◽  
University Hospital ◽  
Aminoglycoside Resistance ◽  
Single Clone ◽  
Aminoglycoside Resistance Genes ◽  
High Level ◽  
Level Resistance

Among 145Enterococcus faecalisisolates recovered during a 15-month period (April 1997-June 1998) in AHEPA University Hospital, Thessaloniki, Greece, 94 (65%) exhibited high-level resistance to gentamicin or streptomycin and 61 (42%) to both aminoglycosides; 73% of the high-level aminoglycoside-resistantE faecalisisolates belonged to a single clone carrying the geneaac(6')-Ie-aph(2”)-Ia. These findings differ from those of other regions, where high-level aminoglycoside-resistance genes are dispersed into genetically unrelated strains.

Epidemiology of Clostridium difficile infection: results of a hospital-based study in Krakow, Poland

2015 ◽  
Vol 143 (15) ◽  
pp. 3235-3243 ◽  
Author(s):  
J. CZEPIEL ◽  
J. KĘDZIERSKA ◽  
G. BIESIADA ◽  
M. BIRCZYŃSKA ◽  
W. PERUCKI ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Fold Increase ◽  
Time Span ◽  
University Hospital ◽  
Posterior Stabilization ◽  
The Past ◽  
Risk Of Mortality ◽  
Crude Mortality ◽  
Medical Wards ◽  
The University

SUMMARYOver the past two decades Clostridium difficile infection (CDI) has appeared as a major public health threat. We performed a retrospective study based on the records of patients hospitalized for CDI at the University Hospital in Krakow, Poland, between 2008 and 2014. In the study period, CDI occurred in 1009 individuals. There were 790 (78%) individuals who developed infection only once, whereas 219 (22%) developed infection more than once. The percentage of deaths within 14 days of CDI confirmation was 2·4%, with a mean age of 74·2 ± 15·9 years. Crude mortality was 12·9% in medical wards, 5·6% for surgical wards and 27·7% in the ICU setting. The time span between diagnosis and death was 5·1 days on average. Between 2008 and 2012 a 6·5-fold increase of CDI frequency with a posterior stabilization and even reduction in 2013 and 2014 was observed. According to the data analysed, 2/3 patients in our population developed CDI during their hospitalization even though they were admitted for different reasons. Medical wards pose a significantly higher risk of CDI than the surgical ones. Age is a risk factor for CDI recurrence. In the case of patients who died, death occurred shortly after diagnosis. The first CDI episode poses much higher risk of mortality than the consecutive ones.

scholarly journals Outbreak of Infections Caused by Pseudomonas aeruginosa Producing VIM-1 Carbapenemase in Greece

2000 ◽  
Vol 38 (3) ◽  
pp. 1290-1292 ◽  
Author(s):  
Athanassios Tsakris ◽  
Spyros Pournaras ◽  
Neil Woodford ◽  
Marie-France I. Palepou ◽  
Gioia S. Babini ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
University Hospital ◽  
Pulsed Field ◽  
Serotype O ◽  
Class B ◽  
High Level ◽  
Level Resistance

Resistance to imipenem and meropenem was observed in 211 (16.5%) isolates of Pseudomonas aeruginosa recovered in a Greek university hospital during 1996 to 1998. In six isolates selected from throughout this period, high-level resistance to both carbapenems (MICs ≥ 128 μg/ml) was associated with production of the class B β-lactamase VIM-1. bla VIM-bearing isolates belonged to serotype O:12 and were indistinguishable by pulsed-field gel electrophoresis.

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