What action should be taken to prevent spread of vancomycin resistant enterococci in European hospitals?
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Biomedical subjects
Publications and source records attributed to Jan Verhoef.
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In Alzheimer's disease (AD), chemotaxis might be responsible for attracting glial cells towards the neuritic plaque. Using primary monocyte-derived macrophages and primary adult astrocytes as a model, amyloid-beta (Abeta) (1-42) was able to stimulate the production, as measured by RT-PCR, of MIP-1alpha and MIP-1beta mRNA in macrophages and MCP-1 in astrocytes. Cocultures showed in unstimulated as well as in Abeta-stimulated cells an increase in MIP-1alpha, MIP-1beta and MCP-1 mRNA. ELISAs of supernatant samples of stimulated macrophages and astrocytes also showed an increase in MIP-1alpha and MIP-1beta in macrophages and MCP-1 in astrocytes. Stimulated cocultures showed an increase in MIP-1alpha, MIP-1beta and MCP-1 protein levels in contrast to unstimulated cocultures.
The susceptibility of nasopharyngeal Streptococcus pneumoniae to eight antibiotics was studied in 482 children under 5 years of age with community-acquired pneumonia and in 429 healthy pneumococci carriers in Fortaleza, Brazil. Serotyping of strains with pooled and type-specific antisera was also performed. Overall, S. pneumoniae was isolated from 499/911 (55%) children. The carriage rate in children attending day-care centres was higher (72%) than in children with pneumonia (50%) (P<0.001). MIC determination in 441 strains revealed 45% to be intermediate penicillin-resistant and 4% high penicillin-resistant strains. Resistance rates to co-trimoxazole and erythromycin were 42 and 23%, respectively. Serotyping of 211 penicillin-resistant and 58 randomly selected penicillin-susceptible isolates showed that 78% of the strains belonged to paediatric serogroups 6, 14, 19 and 23 (86% of the penicillin-resistant and 51% of the penicillin-susceptible strains, P=0.001). Resistance rates of S. pneumoniae to penicillin and co-trimoxazole in Fortaleza were higher than previously reported in Brazil and associated with paediatric serogroups 6, 14, 19 and 23.
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BACKGROUND AND OBJECTIVE: In the Netherlands, the prevalence of methicillin resistance among Staphylococcus aureus isolates has been kept to less than 1% by using active screening programs and isolation. At the University Medical Center Utrecht (UMCU), an active screening program for methicillin-resistant S. aureus (MRSA) in the surgical intensive care unit (ICU) was implemented in 1986. Between 1992 and 2001, only 6 patients with MRSA were admitted to the surgical ICU. However, 4 of these 6 strains were able to spread to 23 other patients and 15 healthcare workers (HCWs). We were surprised by the epidemic behavior of these strains and wondered whether this was exceptional for S. aureus or whether methicillin-susceptible S. aureus (MSSA) was also spreading in the ICU. DESIGN: A 2-month, prospective, observational study to investigate the incidence and spread of MSSA in the surgical ICU of UMCU and historical data collected during a 10-year period regarding MRSA. SETTING: A 10-bed surgical ICU in a 1,042-bed teaching hospital. RESULTS: Weekly swabs revealed the presence of MSSA in 11 (24%) of 45 patients and 16 (22%) of 72 HCWs. Of all 4,105 patient-HCW contacts, there were only 21 episodes in which both the patient and the HCW were found to carry MSSA. With the use of pulsed-field gel electrophoresis, no identical strains could be identified. CONCLUSION: In our surgical ICU, MRSA seems to spread more easily than MSSA, probably because of selection under antibiotic pressure or a still unknown intrinsic factor within MRSA.
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Seventy-four isolates of multiresistant Escherichia coli and Klebsiella spp. recovered during a 3-year period and 17 control strains with genotypically identified beta-lactamases were tested for the production of extended-spectrum beta-lactamases (ESBLs) by using the Etest and the VITEK 1, VITEK 2, and Phoenix automated instruments. The use of the Etest was evaluated by investigating its accuracy in detecting the ESBLs of the control strains and by comparing interpretation results of laboratory technicians and experts. The accuracy of the Etest was 94%. With the Etest as the reference for the clinical strains and the genotype as the reference for the control strains, the automated instruments detected the ESBLs with accuracies of 78% (VITEK 2), 83% (VITEK 1), and 89% (Phoenix). No significant difference between the systems with regard to the control strains was detected. The VITEK 2 did, however, perform less well than the Phoenix (P = 0.03) on the collection of clinical isolates, mainly because of its high percentage of indeterminate test results (11%). No significant difference between the performances of the VITEK 1 and either the VITEK 2 or the Phoenix was found. However, because of its associated BDXpert system the Phoenix showed the best performance. The Etest was found to be an accurate test but was limited by its indeterminate results (4%), its inability to differentiate between K1 hyperproduction and ESBLs, questionable guidelines concerning mutants inside the inhibition zones, and the inability of the technicians to recognize subtle zone deformations.
Of the 419 Moraxella catarrhalis isolates collected during the 1997-1999 European SENTRY surveillance study, 385 (92%) were beta-lactamase positive. Twenty-two (5.7%) produced BRO-2 beta-lactamase. Twenty-one new mutations were found in the putative promoter region of the bro genes. Nineteen percent of all isolates tested were complement sensitive. Resistance to beta-lactams is not linked to the phylogenetic lineages associated with susceptibility to complement.
We evaluated two new automated identification systems, the BD Phoenix (Becton Dickinson) and the VITEK 2 (bioMérieux), for identification of isolates of the Burkholderia cepacia complex (BCC). The test sample included 42 isolates of the highly virulent and epidemic genomovar III, 45 isolates of B. multivorans, and 47 isolates of other members of the BCC. Rates of correct identification by the BD Phoenix and VITEK 2 were similar when all BCC isolates were considered (50 and 53%, respectively) but differed markedly for genomovar III (71 and 38%; P < 0.01) and for B. multivorans (58 and 89%; P < 0.001). For the BD Phoenix as well as the VITEK 2, taking all 134 isolates of the BCC together, rates of correct identification of clinical isolates (56 and 55%, respectively; n = 85) were higher than those of environmental isolates (21 and 39%, respectively; n = 28). Clinical isolates of genomovar III (n = 27) showed correct identification rates of 81% (BD Phoenix) and 48% (VITEK 2) (P < 0.01). Rates of misidentification for BD Phoenix and VITEK 2 were 9 and 17% for genomovar III, 22 and 7% for B. multivorans, and 36 and 13% for the other BCC members (P < 0.01), respectively. More than half of the isolates misidentified by each instrument were identified as Ralstonia pickettii, Ralstonia paucula (CDC IV C-2 group), Alcaligenes faecalis, Achromobacter spp., or, for the VITEK 2, "various nonfermenters." This study reemphasizes that confirmatory identification of BCC, preferably by molecular methods, is highly recommended.
Vancomycin-resistant Enterococcus faecium (VREF) strains represent an important threat in hospital infections in the United States and are found at high frequencies in both the community and farm animals in Europe. We evaluated automated ribotyping for interlaboratory reproducibility by using the restriction enzymes EcoRI and BamHI and compared ribotyping to both amplification of fragment length polymorphism (AFLP) analysis and multilocus sequence typing (MLST) to assess its discriminatory power and capacity for the identification of epidemiologically important strains. Of 19 (EcoRI) and 16 (BamHI) isolates tested in duplicate in two laboratories, 18 (95%) and 16 (100%), respectively, showed reproducible ribotypes. These high reproducibility rates were obtained only after manual refinement of the automated fingerprint analysis. A group of 49 VREF strains initially selected to represent 32 distinct AFLP types were separated into 28 EcoRI ribotypes, 25 BamHI ribotypes, and 28 sequence types. Ribotyping with EcoRI and BamHI was able to discern the host-specific genogroups recently disclosed by AFLP typing and MLST and to distinguish most strains containing the esp gene, a marker specific for strains causing hospital outbreaks. An expandable ribotype identification library was created. We recommend EcoRI as the enzyme of choice for automated ribotyping of VREF strains. Given the high level of discrimination of VREF strains, the high rate of interlaboratory reproducibility, and the potential for the identification of epidemiologically important genotypes, automated ribotyping appears to be a very valuable approach for characterizing VREF strains.
The number of life-threatening opportunistic fungal infections has shown a dramatic increase. However, the diagnosis of candidemia remains difficult. Nucleic acid amplification assays may improve the detection rate and decrease the time needed for detection and identification of Candida spp. Whole blood samples of patients suspected of having candidemia were analyzed using Nucleic Acid Sequence-Based Amplification (NASBA). Furthermore, aliquots of blood cultures of the patients after 2 days of culturing were tested. Eleven data sets from ten patients in two hospitals were generated. None of the whole blood samples was positive in the NASBA assay. Eight samples were positive in the NASBA assay after two days of culturing, whereas only two additional positive samples were found after longer incubation periods. Thus, a two-day culture step is sufficient to greatly improve the sensitivity of the NASBA assay. The NASBA assay detected Candida RNA in three patients. In one patient, the yeast was not detected by automated blood culturing, in another patient the NASBA assay detected the infection two days earlier than the blood culture system.
It is generally assumed that one of the reasons why diabetics are more susceptible to urinary tract infections than non-diabetics is their 'sweet urine'. However, very little information is available on this subject. Therefore, the growth rates of different Escherichia coli strains were studied in human urine with and without added glucose and with and without a constant pH, and compared with their growth rates in Mueller-Hinton broth (MHB). Eight isolates were used (three from blood cultures from urosepsis patients, two urinary isolates, two faecal isolates and one laboratory strain K12). All isolates grew better in MHB than in urine, but with the exception of the laboratory strain, they had the same growth rate in urine. No significant difference was found between the growth rate in urine from diabetics without glucosuria and that in urine from non-diabetics. The addition of glucose (up to a concentration of 1000 mg/dl) to urine and MHB enhanced the growth rate of all isolates. However, very high concentrations of glucose (up to 10000 mg/dl) in urine and MHB caused a decrease in bacterial growth rate when the urinary pH was not kept constant. The stationary phase was reached later and the final bacterial yield was greater when the urine was made less acidic. As the uropathogenic strains did not grow better in urine than the other isolates, it may be concluded that better growth in urine is not one of the causes of the greater virulence of these strains.
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