OBJECTIVES:New molecular tests for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are being rapidly launched in response to the coronavirus disease 2019 (COVID-19) pandemic. The aim of this study was to evaluate the analytical and clinical performance of the VIASURE SARS-CoV-2 S gene RT-PCR Kit on the BD Max™ system and to compare results with those obtained with the cobas® SARS-CoV-2 test on the cobas® 6800 system.METHODS:For testing the analytical performance, reference material was used. Clinical samples (n = 101) obtained from individuals with symptoms compatible with COVID-19 were studied. Oropharyngeal and nasopharyngeal swabs were collected by using either ESwab™ or UTM™ collection systems.RESULTS:When the analytical performance was evaluated, the sample containing the lowest SARS-CoV-2 concentration tested negative with the VIASURE test whereas results obtained with the cobas® test were found to be concordant with the results expected. Six out of the 101 clinical samples (5.9%) showed an inhibition with the VIASURE test. When analysing the remaining 95 clinical samples, 27 were found to be negative with both assays. Of 68 samples that were positive with the cobas® test, the VIASURE test missed 21 (30.9 %) samples. All of those 21 samples had shown Ct values ≥ 31 with the cobas® 6800 system. None of the samples tested positive with the VIASURE test and negative with the cobas® test.CONCLUSIONS:The VIASURE test was impaired by a lack of sensitivity and a relatively high number of invalid results. When using the VIASURE test for routine testing, a significant number of COVID-19-positive samples would have been missed.

Hintergrund: Penicillinhaltige Antibiotika können als seltene Nebenwirkung eine segmentale, durch toxinbildende (Tilivallin) Klebsiella oxytoca verursachte, Colitis mit blutiger Diarrhoe und starken abdominellen Schmerzen verursachen (Högenauer et al. N Engl J Med. 2006). Üblicherweise heilt diese Colitis nach Absetzen des auslösenden Antibiotikums spontan aus. Ob dies auch bei immunsupprimierten Patienten zutrifft ist unbekannt.

Hintergrund: C. difficile wird bei CED-Patienten häufiger nachgewiesen als bei Personen, die nicht an einer CED erkrankt sind.

Background: A recent European study revealed clarithromycin primary resistance in Helicobacter pylori to be the highest in Austria among all 18 participating countries (36.6%; Megraud F, Gut, 2013). Austria was represented by a single center, it is therefore not feasible to infer from this data on regional resistance patterns. As the rate of clarithromycin resistance is crucial for the choice of the eradication regime, we aimed to evaluate primary and secondary resistance rates of H. pylori in Southern Austria in 10 centers.

BACKGROUND:Procalcitonin (PCT) has previously been proposed as useful marker to rule out bloodstream-infection (BSI). The objective of this study was to evaluate the sensitivity of different PCT cut-offs for prediction of BSI in patients with community (CA)- and hospital-acquired (HA)-BSI. METHODS:A total of 898 patients fulfilling systemic-inflammatory-response-syndrome (SIRS) criteria were enrolled in this prospective cohort study at the Medical University of Graz, Austria. Of those 666 patients had positive blood cultures (282 CA-BSI, 384 HA-BSI, enrolled between January 2011 and December 2012) and 232 negative blood cultures (enrolled between January 2011 and July 2011 at the emergency department). Blood samples for determination of laboratory infection markers (e.g. PCT) were collected simultaneously with blood cultures. RESULTS:Procalcitonin was significantly (p < 0.001) higher in SIRS patients with bacteremia/fungemia than in those without. Receiver operating characteristic curve analysis revealed an area under the curve (AUC) value of 0.675 for PCT (95% CI 0.636-0.714) for differentiating patients with BSI from those without. AUC for IL-6 was 0.558 (95% CI 0.515-0.600). However, even at the lowest cut-off evaluated (i.e. 0.1 ng/ml) PCT failed to predict BSI in 7% (n = 46) of patients. In the group of patients with SIRS and negative blood culture 79% (n = 185) had PCT levels > 0.1. CONCLUSION:Procalcitonin was significantly higher in patients with BSI than in those without and superior to IL-6 and CRP. The clinical importance of this is questionable, because a suitable PCT threshold for excluding BSI was not established. An approach where blood cultures are guided by PCT only can therefore not be recommended.

For many years, extended-spectrum-beta-lactamase (ESBL) producing bacteria were a problem mainly located in medical facilities. Within the last decade however, ESBL-producing bacteria have started spreading into the community and the environment. In this study, ESBL-producing Escherichia coli from sewage sludge were collected, analysed and compared to ESBL-E. coli from human urinary tract infections (UTIs). The dominant ESBL-gene-family in both sample groups was blaCTX-M, which is the most prevalent ESBL-gene-family in human infection. Still, the distribution of ESBL genes and the frequency of additional antibiotic resistances differed in the two sample sets. Nevertheless, phenotyping did not divide isolates of the two sources into separate groups, suggesting similar strains in both sample sets. We speculate that an exchange is taking place between the ESBL E. coli populations in infected humans and sewage sludge, most likely by the entry of ESBL E. coli from UTIs into the sewage system.

Background: Klebsiella oxytoca is an opportunistic pathogen and causes diseases like antibiotic-associated hemorrhagic colitis (AAHC) and nosocomial pneumonia. The latter is thought to be caused by strains originating from the intestine. A subgroup of K. oxytoca isolates produces a non-ribosomal cytotoxic peptide, which induces colitis in AAHC and is absent from other Klebsiella spp. Previous attempts to assess the genetic relatedness of K. oxytoca isolates using typing methods such as pulsed field gel electrophoresis were not successful in discriminating different subtypes. To move forward we established a novel multilocus sequence typing (MLST) for K. oxytoca.

This study determined the genetic background of virulence and resistance genes of MRSA ST398 in Austria. From 2004 up to 2008 a total of 41 human isolates of MRSA ST398 were investigated for virulence and resistance gene patterns using DNA microarray chip analysis. Highly similar virulence gene profiles were found in 29 (70·7%) of the isolates but genes encoding Panton-Valentine leukocidin, enterotoxins, or toxic shock syndrome toxin were not detected. Genes conferring resistance to tetracycline and erythromycin-lincosamide were common as all but one of the isolates exhibited tetM and/or tetK, which are involved in tetracycline resistance, and 12 (29·9%) were positive for ermC, conferring resistance to erythromycin/lincosamide. SplitsTree analysis showed that 40 isolates were closely related. Changes in virulence and resistance gene patterns were minimal over the observed time period.

We report the emergence of carbapenem-resistant Enterobacteriaceae in Austria. Over a 10-year period, carbapenem-resistant Enterobacteriaceae isolates were obtained from 13 hospitalized patients, with the first isolation in the year 2005 and a remarkable increase in the number of involved patients in 2010. Carbapenem-resistant Enterobacteriaceae comprise eight Klebsiella pneumoniae isolates, four Klebsiella oxytoca isolates, and one Escherichia coli isolate. The detected carbapenemases were the metallo-β-lactamases New Delhi β-lactamase, VIM and IMP, and the serin-β-lactamase Klebsiella pneumoniae carbapenemase.

Objective: Rapid and reliable diagnosis of genetic relatedness of clinical isolates in microbiologic laboratory is essential in case of nosocomial outbreak investigation. Most molecular techniques used to type microorganisms are technically demanding and time consuming. Currently repetitive-sequence-based PCR (rep-PCR) technique has been adapted to an automated format on the DiversiLab system (bioMerieux, Marcy l'Etoile, France). Aim of this study was to compare the performance of the DiversiLab system to that of pulsed-field gel electrophoresis (PFGE) in nosocomial outbreaks.Methods: 122 clinical isolates (28 Methicillin-resistant Staphylococcus aureus (MRSA), 26 Acinetobacter baumannii, 45 extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae and 13 ESBL-producing Klebsiella oxytoca) were investigated. 70 isolates originated from six well-documented outbreaks, 52 were non-outbreak isolates.Results: Concordant results for identification of outbreak and non-outbreak MRSA, A. baumannii and ESBL-producing K. pneumoniae strains were achieved with both methods. In the outbreak of ESBL-producing K. oxytoca automated rep-PCR was slightly more discriminatory than PFGE. Rep-PCR identified investigated ESBL-producing K. oxytoca outbreak-strains as indistinguishable or closely related, showing similarity of >90%, while PFGE identified these strains as indistinguishable.Conclusion: Automated rep-PCR assays on the DiversiLab system were used for MRSA, A. baumannii and for the first time ESBL-producing Klebsiella spp. and proved as a rapid and reliable method for molecular analysis of nosocomial outbreaks. (C) 2009 The British Infection Society. Published by Elsevier Ltd. All rights reserved.

Einleitung: Klebsiella oxytoca wurde kürzlich als Ursache der Antibiotika assoziierten hämorrhagischen Kolitis (AAHC) nachgewiesen welche vor allem durch Penicilline ausgelöst wird. Die genaue Pathogenese wie K. oxytoca die Kolitis verursacht ist unklar. Histopathologisch ähnelt die AAHC anderen Formen einer Toxin-induzierten Kolitis. Bei einigen K. oxytoca Isolaten von AAHC-Patienten konnte eine Toxinproduktion in einer Zellkultur nachgewiesen werden. Die Fragestellung der aktuellen Studie war ob diese Toxinproduktion nur bei speziellen K. oxytoca Stämme die mit einer Kolitis assoziiert sind auftritt. Methodik: Der Zytotoxineffekt von K. oxytoca wurde in einem Zellkulturassay untersucht. Bei diesem wurden eukaryotische Zelllinien mit bakteriellem Überstand behandelt und das Überleben der Zelllinien detektiert. Insgesamt wurden 121 Isolate (K. oxytoca sowie andere Klebsiella spp.) auf Toxinproduktion getestet. Die Isolate wurden aus dem Stuhl von AAHC Patienten, Patienten mit Kolitis anderer Genese oder Gesunden isoliert. Weiters wurden Stämme von Infektionen in anderen Körperregionen (Harntrakt, Respirationstrakt, Haut und Blut) getestet. Ergebnisse: 69% der K. oxytoca Isolate von AAHC Patienten zeigten eine Zytotoxinproduktion. Bei einzelnen AAHC Patienten wurden sowohl Toxin positive als auch Toxin negative Stämme im selben Patienten gefunden. Andere K. oxytoca Stuhlisolate zeigten auch häufig eine Toxinproduktion: Kolitis anderer Genese: 57%; gesunde Individuen: 46%. Keines der K. oxytoca Isolate von Patienten mit Harnwegs- oder respiratorischen Infektion war Toxin positiv. Nur 15% der Isolate aus Blutkulturen waren Toxin positiv. Alle getesteten Stämme von K. pneumoniae und anderen Klebsiella spp. zeigten keine Toxinproduktion.

Antibiotic resistance in Streptococcus pneumoniae has increased worldwide but varies within geographical regions. We conducted a retrospective analysis of resistance in S. pneumoniae over a 12-year period to assess local and temporal trends in antibacterial resistance. From 1997 to 2008, a total of 1814 non-duplicate S. pneumoniae isolates were identified at the Institute of Hygiene, Microbiology and Environmental Medicine, Medical University of Graz, Austria. Antibiotic resistance was determined by the Clinical and Laboratory Standards Institute (CLSI) disk diffusion test. For penicillin, the minimum inhibitory concentration was determined by Etest. Susceptibility was defined according to CLSI interpretive criteria. For penicillin, resistance rates were consistently low at 0.2% over the 12-year study period. An increase in resistance was remarkable for erythromycin (3.5% in 1997; 14.7% in 2008), clindamycin (1.8% in 1997; 10.6% in 2008) and tetracycline (1.8% in 2000; 11.0% in 2008). For trimethoprim/sulfamethoxazole, resistance increased slightly to 9.2% in 2008. Quinolones showed a low resistance rate of 0.2% that persisted over the whole study period. In contrast to previously published national data, resistance to penicillin was observed to remain at a remarkably low and constant level. Although international surveillance programmes have set up sustainable and interlinked data networks, our results suggest that regional surveillance may still be needed as decision support for appropriate empirical antibiotic therapy in the local health setting.