The aetiology of acute exacerbations of chronic obstructive pulmonary disease (COPD) is heterogeneous and still under discussion. Serological studies have suggested that Mycoplasma pneumoniae, Chlamydia pneumoniae and Legionella pneumophila may play a role in acute exacerbations of COPD.The presence of these atypical pathogens in sputum samples was investigated in patients with stable COPD and with acute exacerbations of COPD using real-time PCR. The present study was part of a randomised, double-blind, single-centre study and a total of 248 sputum samples from 104 COPD patients were included. In total, 122 samples obtained during stable disease (stablestate sputa) and 126 samples obtained during acute exacerbations of COPD (exacerbation sputa) were tested.Of the 122 stable-state sputa, all samples were negative for M. pneumoniae and C. pneumoniae DNA, whereas one sample was positive for Legionella non-pneumophila DNA. Of the 126 exacerbation sputa, all samples were negative for M. pneumoniae and C. pneumoniae DNA, whereas one sample was positive for Legionella non-pneumophila DNA.The possible relationship between the presence of atypical pathogens and the aetiology of acute exacerbations in chronic obstructive pulmonary disease was investigated in patients with stable disease and in those with acute exacerbations using real-time PCR. No indication was found of a role for Legionella spp., Chlamydia pneumoniae or Mycoplasma pneumoniae in stable, moderately severe chronic obstructive pulmonary disease and in its exacerbations.
Inter-and intralaboratory inconsistencies in detection rates of Chlamydia pneumoniae in vascular specimens have been demonstrated. In this study, 66 vascular tissue specimens from 66 patients with vascular disease were tested by three PCR assays: a 16S PCR-based reverse line blot (RLB) assay, a single-step PCR, and a nested PCR. Also, we explored the impacts of different DNA polymerase enzymes on the results based on gel electrophoresis and hybridization. The PCR results by gel electrophoresis in the single-step PCR depended on which DNA polymerase was used. All samples were negative with AmpliTaq Gold DNA polymerase, and 54.5% (36 of 66) were positive with the conventional Taq DNA polymerase. All samples were negative after hybridization with a C. pneumoniae-specific probe. In the nested PCR, all specimens were negative by gel electrophoresis and after hybridization. The RLB assay failed to detect C. pneumoniae in any specimen; however, 20 specimens were Chlamydia sp. positive. The sequence analysis of six of these samples demonstrated Chlamydialike organisms. RLB detected Chlamydia sp. DNA in water and in the elution buffer after passage of the Qiagen columns (11 of 40). This study identified factors that may influence the detection of C. pneumoniae DNA in vascular tissues and consequently bias the perception of a link between C. pneumoniae and vascular diseases. The following are strongly recommended: to use DNA polymerases that have to be activated, to decontaminate with dUTP-uracil-DNA glycosylase, to hybridize with specific probes, to include sufficient controls, and to use molecular grade water.
Background We describe the control of a large VRE outbreak in a Dutch general hospital in 2014–2017. Methods After the outbreak was identified, a screening policy consisting of a single rectal swab culture (with enrichment broth) to identify VRE carriage in high risk patients, was implemented. In addition to screening, measures to improve compliance with standard infection control precautions and enhanced environmental cleaning were implemented to control the outbreak. Results Between September, 2014 and February, 2017, 140 patients were identified to be colonised by vanA mediated vancomycin-resistant Enterococcus faecium (VREfm). Three of these patients developed bacteraemia. AFLP typing showed that the outbreak was caused by a single clone. Extensive environmental contamination was found in multiple wards. Within nine months after the detection of the outbreak no new VRE cases were detected. Conclusion We implemented a control strategy based on targeted screening and isolation in combination with implementation of general precautions and environmental cleaning. The strategy was less stringent than the Dutch national guideline for VRE control. This strategy successfully controlled the outbreak, while it was associated with a reduction in the number of isolation days and the number of cultures taken.
Background:The objective of this study was to determine the correlation between adenosine triphosphate (ATP) measurements and microbial contamination using a standardized method. Methods: ATP measurements and aerobic colony counts (ACC’s) were conducted on 10 pre-defined fomites in a hospital and nursing home setting. Per fomite two ATP measurements and two agar plate measurements were conducted, each measurement was conducted on a 25 cm2 surface. Both measurements were compared and analyzed for correlation. Results: In total 200 paired measurements were conducted, 200 ATP measurements and 200 ACC’s. The mean of all ATP measurements tested on the same surface was calculated, as was for all 200 ACC’s. There was a strong correlation between the mean of two ATP measurements on two different sites on the same fomite (R=0.800, p<0.001) as well as between two ACC measurements on the same fomite (R=0.667, p<0.001). A much weaker correlation was found between RLU values and ACC’s (R=0.244, p<0.001). Conclusions: Reproducibility of ATP measurements and ACC’s on the same fomite was good. However, the correlation between RLU values and ACC’s on hospital surfaces was much lower. This may be explained by the wide variety of biological material that is measured with ATP, of which the bacterial load is only one of many components. ATP measurement can be used to give a quantifiable outcome for the rating of cleanliness in health care facilities, however the results cannot be translated into the level of microbial contamination.
Background & objective: The objective of this study was to determine the effect from feedback of ATP measurements on environmental contamination within hospitals in the Dutch/Belgian border area. Setting: Standardized ATP measurements were conducted in 9 hospitals on pre-defined fomites. Four different fomite groups were defined: medical devices, patient bound materials, ward bound materials and sanitary items. ATP results were reported in relative light units (RLU), RLU >1000 was considered as “not clean.” Two rounds of ATP measurements were conducted. After the first round of ATP measurements, results were provided to the wards. There were no structured cleaning interventions implemented. The second round of ATP measurements was performed one year later. The amount of surface contamination before and after the feedback were compared. Results: In total 1923 ATP measurements were performed. Before feedback 960 ATP measurements were conducted and after feedback 963. The overall median reduction in RLU was 381 (p<0.001), from 568 before feedback to 187 afterwards. In each hospital there was a reduction of the median RLU after feedback. Conclusion/discussion: Substantial reductions in RLU values were found after feedback of ATP measurements. Feedback of ATP measurements was associated with a major reduction of surface contamination in hospitals.
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