How reliable are test results from 17 laboratories on the basis of EN 1500 for a hand rub based on 80% (w/w)?

INTRODUCTION: A ring trial organized by the Association of Applied Hygiene (VAH) on the bactericidal efficacy of an ethanol-based hand rub was carried out in 17 laboratories according to EN 1500 with the aim of describing the variability of test results and fulfilment of the methodological acceptance criteria. METHOD: As a test product, a hand rub based on 80% ethanol (w/w) was tested in comparison with the reference alcohol (60% iso-propanol, v/v) in a crossover design. After pre-washing and contamination following the norm, hands were treated either with the reference alcohol (2 × 3 mL for 2 × 30 s) or the test product (3 mL in 30 s). Post-decontamination values were determined immediately after the rub-in period. Validated neutralizers were used. The arithmetic means of all individual log10 pre-values, post-values and reduction values were calculated per laboratory. Non-inferiority was assumed when the Hodges-Lehmann 97.5% confidence limit was <0.6 in comparison with the reference. A z-score was calculated to determine the laboratory performance. RESULTS: Two laboratories did not meet the acceptance criteria and were excluded from the analysis. The bactericidal efficacy of the test product was non-inferior to the reference product in four laboratories and not non-inferior in 11 laboratories. The z-score for the Hodges-Lehmann 97.5% confidence limit indicated a satisfactory performance in all laboratories. CONCLUSION: We consider the EN 1500 test method to be robust in terms of the variability of test results. For products of borderline efficacy, the evaluation should be based on more than one test.

Failure of sodium hypochlorite to meet the EN 1500 efficacy requirement for hygienic hand disinfection.

This study evaluated the bactericidal efficacy of two alternative hand antiseptics, based on sodium hypochlorite or sodium hypochlorite and hypochlorous acid, compared with isopropanol on the hands of volunteers artificially contaminated with Escherichia coli using EN 1500. The reference alcohol was applied according to the norm, and the study formulations were used as in common practice (3 mL for 30 s). The products showed mean log10 reductions of 1.63 and 1.89, both of which were inferior to the reference treatment (4.78). Due to the failure to achieve sufficient bactericidal efficacy within 30 s, sodium hypochlorite (0.05-0.06%) should not be considered for hand disinfection.

Efficacy of five 'sporicidal' surface disinfectants against Clostridioides difficile spores in suspension tests and 4-field tests.

BACKGROUND: A sporicidal surface disinfection is recommended both for the outbreak and the endemic setting but a comparative evaluation on the efficacy of 'sporicidal' surface disinfectants using suspension tests and 4-field tests has not been performed. AIM: To determine the efficacy of five 'sporicidal' surface disinfectants (three ready-to-use wipes (A, B, E), two concentrates (C, D) based on peroxides or aldehydes against C. difficile spores. METHODS: The efficacy was determined under clean conditions using a suspension test and the 4-field test. Each test was performed in duplicate in two separate laboratories. Wipes were wrung to collect the solution for the suspension tests. RESULTS: Product A (peracetic acid; 5 min), product C (peracetic acid; 2% solution in 15 min or 1% solution in 30 min) and product D (peracetic acid; only 2% solution in 15 min) were effective with at least a 4 log10-reduction of C. difficile spores in suspension and on surfaces. Product B (hydrogen peroxide) was not effective in suspension (0.9 log10 after 15 min; 3.2 log10 after 1 h) and on surfaces (2.8 log10 after 15 and 60 min). Product E based on glutaraldehyde, (ethylendioxy)dimethanol and DDAC demonstrated 0.9 log10 after 4 h in suspension and 4.5 log10 after 4 h on surfaces. CONCLUSIONS: Not all surface disinfectants with a sporicidal claim were effective against C. difficile spores in standardized suspension tests and in the 4-field test. In clinical practice preference should be given to products that reliably pass the efficacy criteria of both types of tests.

Interlaboratory reproducibility of a test method following 4-field test methodology to evaluate the susceptibility of Clostridium difficile spores.

BACKGROUND: Sporicidal surface disinfection is recommended to control transmission of Clostridium difficile in healthcare facilities. EN 17126 provides a method to determine the sporicidal activity in suspension and has been approved as a European standard. In addition, a sporicidal surface test has been proposed. AIM: To determine the interlaboratory reproducibility of a test method for evaluating the susceptibility of a C. difficile spore preparation to a biocidal formulation following the 4-field test (EN 16615 methodology). METHODS: Nine laboratories participated. C. difficile NCTC 13366 spores were used. Glutaraldehyde (1% and 6%; 15 min) and peracetic acid (PAA; 0.01% and 0.04%; 15 min) were used to determine the spores' susceptibility in suspension in triplicate. FINDINGS: One-percent glutaraldehyde revealed a mean decimal log10 reduction of 1.03 with variable results in the nine laboratories (0.37-1.49) and a reproducibility of 0.38. The effect of 6% glutaraldehyde was stronger (mean: 2.05; range: 0.96-4.29; reproducibility: 0.86). PAA revealed similar results. An exemplary biocidal formulation based on 5% PAA was used at 0.5% (non-effective concentration) and 4% (effective concentration) to determine the sporicidal efficacy (4-field test) under clean conditions in triplicate with a contact time of 15 min. When used at 0.5% it demonstrated an overall log10 reduction of 2.68 (range: 2.35-3.57) and at 4% of 4.61 (range: 3.82-5.71). The residual contamination on the three primarily uncontaminated test fields was <50 cfu/25 cm2 in one out of nine laboratories (0.5%) and in seven out of nine laboratories (4%). CONCLUSION: The interlaboratory reproducibility seems to be robust.

Isopropanol at 60% and at 70% are effective against 'isopropanol-tolerant' Enterococcus faecium.

The bactericidal activity of isopropanol was determined against Enterococcus faecium ATCC 6057, ST 796 (isopropanol-tolerant strain) and Enterococcus hirae ATCC 10541 (EN 13727). Isopropanol at 60% and 70% were effective (≥5.38 log10-reduction) in 15 s against all strains but 23% isopropanol was not (<0.99 log10-reduction in ≤15 min). Isopropanol at 70% was tested against E. faecium in the four-field test. Eight millilitres was not effective enough in 1 min (<5 log10-reduction), whilst 16 mL was effective (≥5.85 log10-reduction). Healthcare workers can be reassured that 60% and 70% isopropanol with an appropriate volume are effective against E. faecium.

Variables That Best Differentiate In-Patient Acute Stroke from Stroke-Mimics with Acute Neurological Deficits.

Introduction. Strokes and stroke-mimics have been extensively studied in the emergency department setting. Although in-hospital strokes are less studied in comparison to strokes in the emergency department, they are a source of significant direct and indirect costs. Differentiating in-hospital strokes from stroke-mimics is important. Thus, our study aimed to identify variables that can differentiate in-hospital strokes from stroke-mimics. Methods. We present here a retrospective analysis of 93 patients over a one-year period (2009 to 2010), who were evaluated for a concern of in-hospital strokes. Results. About two-thirds (57) of these patients were determined to have a stroke, and the remaining (36) were stroke-mimics. Patients with in-hospital strokes were more likely to be obese (p = 0.03), have been admitted to the cardiology service (p = 0.01), have atrial fibrillation (p = 0.03), have a weak hand or hemiparesis (p = 0.03), and have a prior history of stroke (p = 0.05), whereas, when the consults were called for "altered mental status" but no other deficits (p < 0.0001), it is likely a stroke-mimic. Conclusion. This study demonstrates that in-hospital strokes are a common occurrence, and knowing the variables can aid in their timely diagnosis and treatment.

Analysis of the oligomeric state and transactivation potential of TAp73α.

The proteins p73 and p63 are members of the p53 protein family and are involved in important developmental processes. Their high sequence identity with the tumor suppressor p53 has suggested that they act as tumor suppressors as well. While p63 has a crucial role in the maintenance of epithelial stem cells and in the quality control of oocytes without a clear role as a tumor suppressor, p73's tumor suppressor activity is well documented. In a recent study we have shown that the transcriptional activity of TAp63α, the isoform responsible for the quality control in oocytes, is regulated by its oligomeric state. The protein forms an inactive, dimeric and compact conformation in resting oocytes, while the detection of DNA damage leads to the formation of an active, tetrameric and open conformation. p73 shows a high sequence identity to p63, including those domains that are crucial in stabilizing its inactive state, thus suggesting that p73's activity might be regulated by its oligomeric state as well. Here, we have investigated the oligomeric state of TAp73α by size exclusion chromatography and detailed domain interaction mapping, and show that in contrast to p63, TAp73α is a constitutive open tetramer. However, its transactivation potential depends on the cellular background and the promoter context. These results imply that the regulation of p73's transcriptional activity might be more closely related to p53 than to p63.

Verification of the efficiency of chemical disinfection and sanitation measures in in-building distribution systems.

Previous investigations of biofilms, generated in a silicone tube model have shown that the number of colony forming units (CFU) can reach 10(7)/cm(2), the total cell count (TCC) of microorganisms can be up to 10(8)cells/cm(2). The present study focuses on the situation in in-building distribution systems. Different chemical disinfectants were tested for their efficacy on drinking water biofilms in silicone tubes: free chlorine (electrochemically activated), chlorine dioxide, hydrogen peroxide (H(2)O(2)), silver, and fruit acids. With regard to the widely differing manufacturers' instructions for the usage of their disinfectants three different variations of the silicone tube model were developed to simulate practical use conditions. First the continuous treatment, second the intermittent treatment, third the efficacy of external disinfection treatment and the monitoring for possible biofilm formation with the Hygiene-Monitor. The working experience showed that it is important to know how to handle the individual disinfectants. Every active ingredient has its own optimal application concerning its concentration, exposure time, physical parameters like pH, temperature or redox potential. When used correctly all products tested were able to reduce the CFU to a value below the detection limit. Most of the active ingredients could not significantly reduce the TCC/cm(2), which means that viable microorganisms may still be present in the system. Thus the question arises what happened with these cells? In some cases SEM pictures of the biofilm matrix after a successful disinfection still showed biofilm residues. According to these results, no general correlation between CFU/cm(2), TCC/cm(2) and the visualised biofilm matrix on the silicone tube surface (SEM) could be demonstrated after a treatment with disinfectants.

Detection of Helicobacter pylori in biofilms by real-time PCR.

Helicobacter pylori is a cause of peptic ulcer disease and a causative agent of gastric cancer. Currently, a possible waterborne route of transmission or a possible survival in drinking water biofilms is discussed. H. pylori, like many other bacterial strains, has the ability to enter the viable but nonculturable state (vbnc) in case of unfavorable conditions. Therefore it is necessary to develop new analysis tools for vbnc bacteria. We established a fast and reliable method to detect H. pylori in drinking water biofilms by quantitative real-time PCR which makes it redundant to use difficult cultivation methods for nonculturable bacteria. With this method it was possible to identify water biofilms as a niche for H. pylori. The real-time PCR analysis targets the ureA subunit of the Helicobacter pylori urea gene which showed high specificity and sensitivity. The quantitative real-time PCR was used to detect H. pylori in biofilms of different age, unspiked and spiked with predetermined levels of cells. The drinking water biofilms were generated in a silicone-tube model. The DNA-sequences for probe and primers showed no cross-homologies to other related bacteria and it was possible to detect less than 10 genomic units of H. pylori. This novel method is a useful tool for a fast screening of drinking water biofilms for H. pylori. The results suggest that drinking water biofilms may act as a reservoir for H. pylori which raises new concerns about the role of biofilms as vectors for pathogens like Helicobacter pylori.

[Risk of infection in the home environment. Plea for a new risk assessment]. / Infektionsrisiken im häuslichen Umfeld. Plädoyer für eine neue Risikobewertung.

Within the last two decades risks posed by infectious diseases outside of hospitals and nursing homes had no high significance in the public perception. The home environment is regarded as a save shelter from infectious risks. In the mean time there have been dramatic socio-demographic, health policy and technological changes which have increased infectious risks outside medical facilities. In Germany up to 1.4 million people with multiple morbidities are nursed at home. Technological changes with the aim to protect the environment, like reduction of water temperature and water volumes in washing processes has lowered the efficacy to control pathogens. Thus it is time to revise the process of risk assessment in which not only aspects of environmental protection but also those of health protection must be taken into account. The article gives an overview of new risks and epidemiological changes and discusses the necessity of a new risk assessment and risk management approach which hopefully will lead to a changing paradigm.

[Hygiene tips for kids. Concept and examples of realisation]. / Hygiene-Tipps für Kids. Konzept und Umsetzungsbeispiele.

Basic hygiene measures aim at promoting and maintaining good health. The necessary routines are most effectively learned and adopted during childhood. These key findings form the basis for the educational programme "Hygiene Tips for Kids" which was initiated at the Department of Hygiene and Public Health at Bonn University under the auspices of WHO Europe in 2003. The programme addresses children, teachers and parents and offers a range of materials and activities tailored to the specific needs of the target groups based on expert advice. The objective is to exert a long-lasting, positive influence on the performance of hygiene routines. Campaigns are usually directed at pre-school, kindergarten and primary school settings, with the public health service personnel playing a crucial role in coordinating and implementing the various activities. Evaluation of a campaign in the federal state of Mecklenburg-Vorpommern has proven a positive effect on hand washing routines in pre-school as well as in the domestic setting. Moreover, Hygiene Tips for Kids has a sustained positive effect on the cooperation and communication between public health authorities and teachers, children and their parents. Teachers implement their own ideas within a framework of standardised information to suit their local needs. A long-term goal would be the obligatory integration of a sensible health education programme in the syllabus of pre-school, kindergarten and primary schools.

[Water as a reservoir for nosocomial infections in health care facilities, prevention and control]. / Wasser als Infektionsquelle in medizinischen Einrichtungen, Prävention und Kontrolle.

New epidemiological and microbiological investigations using molecular typing methods to link patient and environmental strains demonstrate a strong association between water-borne pathogens and nosocomial infections. Avoiding patient exposure to these pathogens results in a decreased incidence of water-borne nosocomial infections. There remains a tremendous potential to reduce hospital acquired infections previously viewed as inevitable and unavoidable through intervention and preventive measures. The characteristics of water application in health care facilities, the vulnerability of patients, the spectrum of relevant pathogens and their ecologic aspects, legal issues and important measures for prevention and control are discussed.

Pretreatment ipsilateral regional cortical blood flow influences vessel recanalization in intra-arterial thrombolysis for MCA occlusion.

BACKGROUND AND PURPOSE: The aim of acute stroke interventions is to achieve recanalization of the target occluded artery. We sought to determine whether pretreatment cortical cerebral blood flow (CBF) was associated with vessel recanalization in patients undergoing intra-arterial therapy. METHODS: This is a retrospective analysis of patients who underwent a quantitative xenon CT blood flow study and were noted to have a documented M1 middle cerebral artery (MCA) or carotid terminus occlusion less than 6 hours from symptom onset between January 1997 and April 2001. Twenty-three patients who underwent intra-arterial thrombolysis were included in the analysis. Univariate and multivariate analyses were performed to determine whether pretherapy CBF was correlated to the likelihood of recanalization. RESULTS: A total of 23 patients were studied in this analysis with a median age of 69 (range 32-81) and median National Institutes of Health Stroke Score of 19 (range, 8-22). Twelve patients (52%) underwent combined intravenous/intra-arterial therapy, and 11 patients (48%) were treated with intra-arterial thrombolytics alone. Successful vessel recanalization (Thrombolysis in Myocardial Infarction classification 2 or 3 flow) occurred in 13 patients (57%). The only variable associated with recanalization in multivariate modeling was mean ipsilateral MCA CBF (odds ratio, 1.25; 95% confidence interval, 1.01-1.54; P = .035). A receiver operating characteristic curve was generated, and a mean ipsilateral MCA CBF threshold of 18 mL/100 g/min was found to be the threshold for successful recanalization. CONCLUSIONS: Our study suggests that patients with higher mean ipsilateral MCA CBF are more likely to recanalize. The threshold for successful revascularization may be 18 mL/100 g/min. Further study is required to determine whether pretreatment CBF is related to recanalization success.

Process-independent quantitative assessment of residual biological contamination of medical devices reprocessed in washer-disinfectors.

A method based on measuring a soil-induced fluorescence intensity response of 1,8-anilinonaphthalene sulfonate at two fixed wavelengths (460 and 510 nm) was used for determining residual contamination on test soil carriers simulating medical devices after passage through a hospital washer-disinfector. The fluorescence response can be satisfactorily calibrated to soil levels as low as approximately 1 microg/L. Practical tests were performed in two hospitals with washer-disinfectors of 3 types and with several chemical or enzymic cleansers-disinfectants. In combination with the previously developed system of standardized test soil carriers simulating both easily and poorly accessible parts of soiled medical devices, the liver-lactose-oil test soil and an efficient sonication procedure for stripping the residual soil off the carriers, this soil detection method permits the detection of very low contamination levels.