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Author(s): Jiri Dresler[sup.1] , Marcela Krutova[sup.2] , Alena Fucikova[sup.3] , Jana Klimentova[sup.3] , Veronika Hruzova[sup.1] , Miloslava Duracova[sup.3] , Katerina Houdkova[sup.1] , Barbora Salovska[sup.1] , Jana Matejkova[sup.2] , Martin Hubalek[sup.4] [...]

Aim: To perform a retrospective analysis of the high occurrence of Clostridium difficile infection in the surgical department of a Czech tertiary care hospital and to identify weaknesses in C. difficile infection (CDI) prevention and control policies. Methods: Clinical and epidemiological data on eleven CDI cases were collected. C. difficile isolates were characterized by capillary electrophoresis ribotyping, multilocus variable tandem repeat analysis (MLVA), gyrA gene fragment sequencing, and erm(B) fragment PCR amplification. Antibiotic susceptibility to metronidazole, vancomycin, ciprofloxacin, moxifloxacin, and clindamycin was tested. Findings: Eleven CDI cases were caused by C. difficile PCR ribotype 001 strains. These strains revealed two different MLVA profiles with 11 tandem repeat differences. All isolates were susceptible to metronidazole and vancomycin and resistant to ciprofloxacin (MIC ≥32 mg/L), moxifloxacin (MIC ≥32 mg/L), and clindamycin (MIC ≥256 mg/L). All isolates revealed amino acid substitution Thr82Ile, in the GyrA and were erm(B) negative. Conclusion: Two fluoroquinolone and clindamycin-resistant C. difficile PCR ribotype 001 strain clusters occurred at one of the surgical departments of a tertiary care hospital. Ineffective decontamination with suboptimal concentration and time of exposure of sporicidal disinfectants may have resulted in C. difficile transmission. [ABSTRACT FROM AUTHOR]

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., (This article is copyright of The Authors, 2016.)

The treatment of infections caused by Clostridium difficile requires a comprehensive approach. The administration of antibiotics of choice (vancomycin or metronidazole) is crucial. The choice of particular agents is based on the severity and course of the disease. Besides antibiotics, alternative approaches continue to be of interest, especially in forms inadequately responding to antibiotic therapy and in case of recurring infections.

In recent years, Clostridium difficile has been among the most important nosocomial pathogens. It causes intestinal infectious diseases detectable by numerous laboratory methods. Given the severity of the diseases, some tests have proved inadequately sensitive. The article summarizes the first experience with the latest kits for early and sensitive detection of toxic strains of Clostridium difficile.

Incidence of Gram-positive infections caused by bacteria resistant to commonly used antibiotics has increased in the last decades. Resistant strains appeared later in the Czech Republic, however their number has been increasing and new antibiotics have to be used. The greatest increase of frequency can be seen in infections caused by methicillin-resistant Staphylococcus aureus and coagulase-negative staphylococci. Vancomycin-resistant enterococci are usually found in hematooncology patients. Curative use of vancomycin is limited due to a narrow spectrum of activity, nephrotoxicity, and limited penetration into tissues (lung) and cerebrospinal fluid. Linezolid is a good option mainly in infections of skin and soft tissues, and it has an evincible advantage over vancomycin in the treatment of nosocomial pneumonia and surgical-site infections. Oral formulations are favourable allowing switch therapy and earlier discharge from hospital.

Background: One of the problems of contemporary medicine is an increasing number of bacterial strains with hazardous phenotypes of resistance. The feared bacterial pathogens include Klebsiella pneumoniae strains producing AmpA extended-spectrum beta-lactamases. The study focused on the molecular biological characteristics of ESBL-positive strains of Klebsiella pneumoniae collected in the Czech Republic., Materials and Methods: Clinical material from patients hospitalized in 16 Czech hospitals in September and October 2004 was used to isolate and determine Klebsiella pneumoniae strains by standard identification procedures. Their susceptibility to antibiotics was tested using a dilution micromethod. A Double-Disk Synergy Test was used for phenotype determination of ESBL production. The blaTEM, blaSHV and blaOXA genes coding ESBL production were demonstrated by PCR. Molecular biological characteristics of ESBL-positive strains utilized the genomic DNA isolation, XbaI restrictase digestion and PFGE differentiation. The acquired restriction maps of individual isolates were compared using GelCompar II software and their relationship was determined., Results: During the monitored period, 913 Klebsiella pneumoniae strains causing clinically detectable diseases were isolated. Of these, 234 (25.6 %) were determined as ESBL-positive strains. The prevalence of ESBL-positive strains was 38.5 % in ICUs and 15.8 % in standard wards. More than 50 % of ESBL-positive isolates were effectively treated only with meropenem (98 %), cefoperazone/sulbactam (61 %) and amikacin (54 %). Conversely, ESBL-negative strains showed high susceptibility to all tested antibiotics (76-99 %). The molecular biological analysis identified 18 clonal types containing 2-6 identical strains. 17 clones usually contained isolates from one hospital and only in one clone strains from two hospitals were identified., Conclusion: Based on the above mentioned results, the prevalence of ESBL-positive strains of Klebsiella pneumoniae in the Czech Republic can be perceived as relatively high, especially in the ICUs. An extensive spread of epidemic clones within Czech hospitals and, to a limited extent, between them can be demonstrated.

We developed a nested PCR assay that detects the recA gene of the Burkholderia cepacia complex in sputum. The product of the first PCR round is also used to identify the genomovar of the pathogen. The protocol achieves high sensitivity and specificity with simple interpretation of genomovar status.