Should we routinely disinfect floors?
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Attitude of Health Personnel↗
Biomedical subjects
J Kluytmans
Publications and source records attributed to J Kluytmans.
Correlation between detection methods of Chlamydia pneumoniae in atherosclerotic and non-atherosclerotic tissues.
Polymerase chain reaction (PCR) and immunohistochemistry (IHC) have been used to detect Chlamydia (C.) pneumoniae in vascular tissues. Discrepancies between the results of these two methods have frequently been reported. However, the correlation between PCR and IHC has not been analyzed yet. This study assesses the correlation between the detection of C. pneumoniae by PCR and IHC in 45 atherosclerotic and 50 non-atherosclerotic tissue specimens. Also, the presence of Mycoplasma (M.) pneumoniae in these 95 specimens was investigated. Correlation was found between the detection of C. pneumoniae by PCR and IHC in the atherosclerotic tissues. Both tests were positive in 10 specimens and negative in 17 specimens (p = 0.003). There was no significant correlation between PCR and IHC in non-atherosclerotic specimens (p = ns). M. pneumoniae was detected, by PCR, in one atherosclerotic specimen.The results show correlation between PCR and IHC in the detection of C. pneumoniae in atherosclerotic tissues, emphasize the association between C. pneumoniae and atherosclerosis, and support the specificity of the association between C. pneumoniae and atherosclerosis.
Adult↗
A European perspective on nosocomial urinary tract infections II. Report on incidence, clinical characteristics and outcome (ESGNI-004 study). European Study Group on Nosocomial Infection.
OBJECTIVES: To estimate the incidence of nosocomially acquired urinary tract infections (NAUTI) in Europe and provide information on the clinical characteristics, underlying conditions, etiology, management and outcome of patients. MATERIALS AND METHODS: We collected clinical information from NAUTI patients with a microbiology report on the named study day. RESULTS: A total of 141 hospitals from 25 European countries participated in the study. Written institutional bladder catheter guidelines were in place in 90.3% of EU hospitals and 55% of non-EU hospitals (P < 0.05). The total number of new NAUTI episodes on the day of the study was 298, representing an incidence of 3.55 episodes/1000 patient-days and an estimated prevalence of 10.65/1000. The five most commonly isolated micro-organisms were Escherichia coli, Enterococcus sp., Candida sp., Klebsiella sp. and Pseudomonas aeruginosa. Patients from non-EU countries were younger, with more severe underlying diseases with a higher incidence of obstructive uropathy/lithiasis. Overall, 22.8% of patients had no 'classic' UTI-predisposing factors. Catheter-associated UTI (CAUTI) was present in 187 patients (62.8%). A closed drainage system was used in only 78.5% of catheterised patients. The indication for bladder catheterisation was not considered adequate in 7.6% of cases and continuation of bladder catheterisation was considered unnecessary in 31.3%. Opening of the closed drainage system was the most frequent major error in catheter management (16.8%). Antimicrobial treatment was not considered adequate in 19.8% of all cases. CONCLUSIONS: The incidence of NAUTI in a large European population is 3.55/1000 patient-days. There is clearly room for improvement in the area of bladder catheterisation, catheter care and medical management of NAUTI. We recommend that European authorities draw up and implement practical and specific guidelines to reduce the incidence of this infection.
Age Factors↗
A European perspective on nosocomial urinary tract infections I. Report on the microbiology workload, etiology and antimicrobial susceptibility (ESGNI-003 study). European Study Group on Nosocomial Infections.
OBJECTIVES: To obtain information on the microbiology workload, etiology and antimicrobial susceptibility of urinary tract infection (UTI) pathogens isolated in European hospitals. MATERIALS AND METHODS: We collected data available in the microbiology units of a large sample of European hospitals regarding the laboratory workload, diagnostic criteria, and etiology and antimicrobial resistance of the urinary isolates collected on one day (the study day). RESULTS: Data were received from a total of 228 hospitals from 29 European countries. The average rate of urine samples cultured per 1000 admissions in 1999 was 324. The criteria to consider a positive urine culture as significant were quite variable; > or =10(4) colony-forming units (CFU)/mL for bacteria or > or =10(3) CFU/mL in the case of yeasts were the most used cut-off points. On the study day, a total of 607 micro-organisms from 522 patients with nosocomial UTI were isolated. The six most commonly isolated micro-organisms were, in decreasing order: Escherichia coli (35.6%), Enterococci (15.8%), Candida (9.4%), Klebsiella (8.3%), Proteus (7.9%) and Pseudomonas aeruginosa (6.9%). Pseudomonas was isolated more frequently in non-EU countries. The study data reveal high rates of antimicrobial resistance in UTI pathogens, especially in non-EU countries, where Pseudomonas aeruginosa presented rates of aminoglycoside resistance as high as 72% to gentamicin, 69.2% to tobramycin and 40% to amikacin. CONCLUSIONS: Nosocomial UTI accounts for an important proportion of the workload in microbiology laboratories. A consensus on the practice and interpretation of urine cultures in Europe is needed. The levels and patterns of resistance of UTI pathogens must be a serious cause for concern and a clear reason for stricter guidelines and regulations in antimicrobial policy.
Anti-Bacterial Agents↗
International multicenter evaluation of latex agglutination tests for identification of Staphylococcus aureus.
A newly marketed rapid agglutination kit for the identification of Staphylococcus aureus, Slidex Staph Plus (bioMérieux), was compared to Staphaurex Plus (Murex Diagnostics) and Pastorex Staph-Plus (Sanofi Diagnostics Pasteur). The study took place in three clinical microbiology laboratories in three different European countries. A total of 892 staphylococcal isolates, including 278 methicillin-sensitive S. aureus (MSSA) isolates, 171 methicillin-resistant S. aureus (MRSA) isolates, and 443 coagulase-negative staphylococcal isolates, were analyzed. The sensitivities (MSSA/MRSA) and specificities, respectively, were 98. 2% (98.9%/97.1%) and 98.9% for Slidex Staph Plus, 98.2% (98.2%/98. 2%) and 96.2% for Staphaurex Plus, and 98.7% (98.6%/98.8%) and 95.7% for Pastorex Staph Plus. The specificity of the Slidex Staph Plus kit was statistically significantly higher than the specificities of Staphaurex Plus and Pastorex Staph-Plus. The Slidex Staph Plus is a very reliable test for the identification of S. aureus.
Coagulase↗
Rapid slide latex agglutination test for detection of methicillin resistance in Staphylococcus aureus.
The MRSA screen test (Denka Seiken Co., Ltd.), a commercially available, rapid (20-min) slide latex agglutination test for the determination of methicillin resistance by detection of PBP 2a in Staphylococcus aureus, was compared with the oxacillin agar screen test and PCR detection of the mecA gene. A total of 563 S. aureus isolates were tested. Two hundred ninety-six of the isolates were methicillin-susceptible isolates from cultures of blood from consecutive patients. Also, 267 methicillin-resistant isolates that comprised 248 different phage types were tested. Methicillin resistance was defined as the presence of the mecA gene. Of the 267 mecA gene-positive isolates, 263 were positive by the MRSA screen test (sensitivity, 98.5%), and all the mecA-gene negative strains were negative by the MRSA screen test (specificity, 100%). The oxacillin agar screen test detected methicillin resistance in 250 of the mecA gene-positive isolates (sensitivity, 93.6%). The sensitivity of the MRSA screen test was statistically significantly higher than the sensitivity of the oxacillin agar screen test (P < 0. 05). The MRSA screen test is a highly sensitive and specific test for the detection of methicillin resistance. Also, it offers results within half an hour and is easy to perform, which makes this test a valuable tool in the ongoing battle against methicillin-resistant S. aureus.
Latex Fixation Tests↗
Reduction of surgical site infections in major surgery by elimination of nasal carriage of Staphylococcus aureus.
Staphylococcus aureus has long been recognized as an important pathogen in human disease. Staphylococcal infections occur regularly in hospital patients and, despite antibiotic therapy, have severe consequences. An increasing number of such infections are caused by methicillin-resistant S. aureus (MRSA) strains, many of which have become multi-resistant to treatment. In an unblinded intervention trial, with historical controls, perioperative nasal carriage of S. aureus was eliminated using mupirocin nasal ointment. A significant reduction in surgical site infection was observed post-intervention in the treated group of patients. No resistant to mupirocin was observed. The results of this study warrant a prospective randomized, placebo-controlled study to confirm the efficacy of mupirocin.
Carrier State↗
Multicenter evaluation of a modified protocol for the RAPIDEC staph system for direct identification of Staphylococcus aureus in blood cultures.
A modified protocol for the RAPIDEC Staph system (bioMérieux, Marcy-l'Etoile, France) for direct identification of Staphylococcus aureus in blood cultures was evaluated in a multicenter study. A total of 129 blood cultures (BACTEC 9000 Blood Culture System; Becton Dickinson Diagnostic Instrument Systems, Sparks, Md.) containing gram-positive cocci in clusters were analyzed by conventional methods and by RAPIDEC Staph in accordance with the manufacturer's protocol and in accordance with a modified protocol. The sensitivity, specificity, and positive and negative predictive values obtained with the manufacturer's protocol were 90.5, 97.7, 95. 0, and 95.5%, respectively, and those obtained with the modified protocol were 100, 96.6, 93.3, and 100%, respectively. The modified protocol for the RAPIDEC Staph is easier to perform than the manufacturer's protocol and is very reliable.
Bacteremia↗
Outbreak of Staphylococcus schleiferi wound infections: strain characterization by randomly amplified polymorphic DNA analysis, PCR ribotyping, conventional ribotyping, and pulsed-field gel electrophoresis.
Within a 1-year period, six surgical-site infections (SSI) caused by Staphylococcus schleiferi were observed in the department of cardiac surgery of Ignatius Hospital, Breda, The Netherlands. Since outbreaks caused by this species of coagulase-negative staphylococci have not been described before, an extensive environmental survey and a case control study were performed in combination with molecular typing of the causative microorganism in order to identify potential sources of infection. Variability, as detected by four different genotyping methods (random amplification of polymorphic DNA [RAPD], conventional and PCR-mediated ribotyping, and pulsed-field gel electrophoresis [PFGE] of DNA macro restriction fragments), appeared to be limited both among the clinical isolates and among several control strains obtained from various unrelated sources. Among unrelated strains, RAPD and PCR-mediated ribotyping identified two types only, whereas seven different types were identified in a relatively concordant manner by conventional ribotyping and PFGE. The latter two procedures proved to be the most useful tools for tracking the epidemiology of S. schleiferi. Four of the outbreak-related strains were identical by both methods, and two isolates showed limited differences. In the search for a potential source of S. schleiferi infection, two slightly different PFGE types were encountered on several occasions in the nose of a single surgeon. These strains were, however, clearly different from the outbreak type. In contrast, S. schleiferi cultures remained negative for two persons identified on the basis of case control analysis. It was demonstrated that SSI caused by S. schleiferi had a clinical impact for patients comparable to that of a wound infection caused by Staphylococcus aureus. This report describes the first well-documented outbreak of S. schleiferi infection. A source of the outbreak was not detected.
Aged↗
Simultaneous persistence of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus in a neonatal ward of a Warsaw hospital.
Fifty-seven methicillin-resistant Staphylococcus aureus (MRSA) isolates from babies (N = 31), carriers amongst health care workers (N = 16; 10% of all staff members) and the environment (N = 10); 39 MSSA isolates, from babies (N = 18), health care workers (N = 5) and environment (N = 16) were analysed. The strains were from the neonatal ward of a teaching hospital in Warsaw and were collected over a period of 16 months (1993/1994). The isolates were characterized by phage-typing, arbitrary-primed polymerase chain reaction (AP PCR), DNA repeat polymorphism within the protein A gene and the resistance pattern to antimicrobial agents. The presence of the mecA gene was determined by PCR. MRSA were classified as heterogeneously resistant to methicillin, susceptible to other antimicrobial agents and, except for three isolates, appeared to be genotypically almost identical. The first example of mupirocin resistant MRSA in Poland was documented. Amongst MSSA isolates, increased variability was seen, however, the persistence of one predominate clone of MSSA was shown. In this particular hospital environment, several different strains of both MRSA and MSSA were capable of maintaining persistent colonization.
Bacteriophage Typing↗
Coagulase and protein A polymorphisms do not contribute to persistence of nasal colonisation by Staphylococcus aureus.
The nasal carriage rate of Staphylococcus aureus was examined in a longitudinal study of 31 healthy Danish volunteers. Each person was classified as persistent (>8 positive cultures from 10 examinations), an intermittent carrier (50-80% positive cultures) or an ocassional carrier (positive cultures on 10-40% of ocassions only). One hundred and twenty strains from these persons were subjected to phage typing and random amplification of polymorphic DNA (RAPD) analysis. Phage and RAPD typing were in close agreement. RAPD confirmed the spread of a particular S. aureus clone (phage type 95) throughout Denmark. However, no common genotype or phenotype characteristics of S. aureus that could separate persistent from intermittent or incidental colonisers were identified. The immunoglobulin binding protein A and the prothrombin binding coagulase protein are both putative S. aureus virulence or defence factors. Analysis of polymorphisms in the variable repeat regions in the genes for these proteins showed no correlation between the number of repeat units and, consequently, the protein structure with the ability of strains to persist in the human nasal mucosa. The amount of protein A, detectable by its IgG binding activity, appeared not to be correlated to persistence of carriage. Thus protein A and coagulase gene polymorphisms do not seem to play a significant role in the propensity of S. aureus to colonise human nasal epithelium. Furthermore, based on the genetic heterogeneity encountered among the S. aureus strains it is suggested that within the current study population, no single clonal lineage of S. aureus has increased capability to colonise the human nasal epithelium.
Adult↗
Nasal carriage of Staphylococcus aureus: epidemiology, underlying mechanisms, and associated risks.
Staphylococcus aureus has long been recognized as an important pathogen in human disease. Due to an increasing number of infections caused by methicillin-resistant S. aureus (MRSA) strains, therapy has become problematic. Therefore, prevention of staphylococcal infections has become more important. Carriage of S. aureus appears to play a key role in the epidemiology and pathogenesis of infection. The ecological niches of S. aureus are the anterior nares. In healthy subjects, over time, three patterns of carriage can be distinguished: about 20% of people are persistent carriers, 60% are intermittent carriers, and approximately 20% almost never carry S. aureus. The molecular basis of the carrier state remains to be elucidated. In patients who repeatedly puncture the skin (e.g., hemodialysis or continuous ambulatory peritoneal dialysis [CAPD] patients and intravenous drug addicts) and patients with human immunodeficiency virus (HIV) infection, increased carriage rates are found. Carriage has been identified as an important risk factor for infection in patients undergoing surgery, those on hemodialysis or CAPD, those with HIV infection and AIDS, those with intravascular devices, and those colonized with MRSA. Elimination of carriage has been found to reduce the infection rates in surgical patients and those on hemodialysis and CAPD. Elimination of carriage appears to be an attractive preventive strategy in patients at risk. Further studies are needed to optimize this strategy and to define the groups at risk.
Bacterial Adhesion↗
Are variable repeats in the spa gene suitable targets for epidemiological studies of methicillin-resistant Staphylococcus aureus strains?
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Genes, Bacterial↗
Molecular epidemiology of apparent outbreak of invasive aspergillosis in a hematology ward.
During a 2-month period, five patients suffering from invasive infections caused by Aspergillus flavus or Aspergillus fumigatus were identified in the Hematology Department of the University Hospital Dijkzigt (Rotterdam, The Netherlands). To study the epidemiological aspects of invasive aspergillosis, strains from these patients and from the hospital environment, isolated during extensive microbiological screening, were subjected to genotyping. A novel DNA extraction technique, involving freezing, grinding, and direct lysis in guanidium isothiocyanate-containing buffers of mycelial material, was applied. DNA isolation was followed by typing by random amplification of polymorphic DNA (RAPD) analysis. This showed that strains isolated from all patients infected with the same fungal species were genotypically distinct, thus providing evidence against the possibility of an ongoing, single-source nosocomial outbreak. Strains could also be differentiated from strains of geographically diverse origins. However, an A. flavus strain from one of the patients was also frequently encountered in the hospital environment. As all environmental strains were collected after this patient had been diagnosed with invasive disease, the epidemiological value of this observation could not be ascertained. Intensive investigations showed no single source of A. flavus or other aspergilli. RAPD genotyping proved that the outbreak of invasive aspergillosis in the hematology ward consisted of a series of unrelated events and was not due to a common source within the hospital. RAPD fingerprinting of aspergilli may greatly facilitate future investigations of the epidemiology of invasive disease caused by these pathogens.
Adult↗
Comparison of five tests for identification of Staphylococcus aureus from clinical samples.
Five different laboratory tests for the identification of Staphylococcus aureus were compared. Analyses of 271 presumptive S. aureus strains, supplemented with 59 well-defined methicillin-resistant S. aureus (MRSA) isolates, were performed. Only the Staphaurex Plus (Murex Diagnostics, Dartford, United Kingdom) and the Pastorex Staphplus (Sanofi, Marnes-La-Coquette, France) tests displayed 100% sensitivity. The observed difference with the free-coagulase test (Bacto coagulase plasma; Difco, Detroit, Mich.), a bound-coagulase (clumping factor) test, and the former Staphaurex test (Murex Diagnostics) was caused mainly by the inability of these three tests to identify some MRSA strains correctly. Among Polish MRSA isolates included in the analysis, a group of free-coagulase-negative S. aureus strains was detected. Genetic typing by random amplification of polymorphic DNA revealed that the strains showing aberrant behavior when the different test results were compared belonged to limited number of S. aureus clones.
Bacterial Typing Techniques↗
Subdural empyema caused by Escherichia coli: hematogenous dissemination to a preexisting chronic subdural hematoma.
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Aged↗
Food-initiated outbreak of methicillin-resistant Staphylococcus aureus analyzed by pheno- and genotyping.
An outbreak of methicillin-resistant Staphylococcus aureus (MRSA) involving 27 patients and 14 health-care workers (HCW) was studied. The outbreak started in the hematology unit of the University Hospital Rotterdam, Dijkzigt, The Netherlands, and spread to the surgical unit. Twenty-one patients (77.8%) developed clinical disease, and five died. Subsequently, MRSA was detected in food and in the throat of one of the HCW who prepared food for hematology patients. Food contaminated by an HCW most likely caused the first case of MRSA septicemia. This route of transmission has not been described before. The outbreak strain was probably transmitted to the surgical unit by a colonized nurse, where it caused an explosive outbreak. Airborne probably transmitted to the surgical unit by a colonized nurse, where it caused an explosive outbreak. Airborne MRSA transmission played an important role in disseminating the organism. The outbreak was controlled within 6 months by intensifying surveillance, temporarily closing the affected wards, treating carriers, and instituting an MRSA ward outside the hospital. Phage typing, insertion sequence probing, protein A gene typing, and DNA fingerprinting by PCR revealed that all outbreak-related isolates were identical. By pulsed-field gel electrophoresis, all but one of the outbreak-related isolates were determined to be identical. Protein A gene typing identified numerous (11) repeat units in all outbreak-related isolates, which supports the suggestion that the outbreak strain may have been more virulent and more transmissible than other MRSA strains. Pheno- and genotyping studies underlined the value of DNA fingerprinting methods for investigation of MRSA epidemiology. Optimal discriminatory power was achieved by combining the results of four genotyping methods.
Base Sequence↗
Multicenter evaluation of arbitrarily primed PCR for typing of Staphylococcus aureus strains.
Fifty-nine isolates of Staphylococcus aureus and a single strain of Staphylococcus intermedius were typed by arbitrarily primed PCR (AP-PCR). To study reproducibility and discriminatory abilities, AP-PCR was carried out in seven laboratories with a standardized amplification protocol, template DNA isolated in a single institution, and a common set of three primers with different resolving powers. The 60 strains could be divided into 16 to 30 different genetic types, depending on the laboratory. This difference in resolution was due to differences in technical procedures (as shown by the deliberate introduction of experimental variables) and/or the interpretation of the DNA fingerprints. However, this did not hamper the epidemiologically correct clustering of related strains. The average number of different genotypes identified exceeded those of the more traditional typing strategies (F. C. Tenover, R. Arbeit, G. Archer, J. Biddle, S. Byrne, R. Goering, G. Hancock, G. A. Hebert, B. Hill, R. Hollis, W. R. Jarvis, B. Kreiswirth, W. Eisner, J. Maslow, L. K. McDougal, J. M. Miller, M. Mulligan, and M. A. Pfaller, J. Clin. Microbiol. 32:407-415, 1994). Comparison of AP-PCR with pulsed-field gel electrophoresis (PFGE) indicated the existence of strains with constant PFGE types but variable AP-PCR types. The reverse (constant AP-PCR and variable PFGE patterns) was also observed. This indicates additional resolution for combined analyses. It is concluded that AP-PCR is well suited for genetic analysis and monitoring of nosocomial spreading of staphylococci. The interlaboratory reproducibility of DNA-banding patterns and the intralaboratory standardization need improvement.
Bacterial Typing Techniques↗