Candidemia poses a major threat to ICU patients and is routinely diagnosed by blood culture, which is known for its low sensitivity and long turnaround times. We compared the performance of a novel, Candida-specific multiplex real-time PCR assay (Fungiplex^® Candida IVD Real-Time PCR Kit) with blood culture and another established diagnostic real-time PCR assay (LightCycler SeptiFast Test) with respect to Candida detection from whole blood samples. Clinical samples from 58 patients were analyzed by standard blood culture (BC) and simultaneously tested with the Fungiplex Candida PCR (FP) and the SeptiFast test (SF) for molecular detection of Candida spp. Compared to BC, the FP test showed high diagnostic power, with a sensitivity of 100% and a specificity of 94.1%. Overall diagnostic accuracy reached 94.6%. Using SF, we found a sensitivity of 60%, a specificity of 96.1%, and an overall diagnostic accuracy of 92.9%. The Fungiplex Candida PCR has shown good sensitivity and specificity on clinical samples of high-risk patients for direct detection of Candida species in whole blood samples. Together with conventional diagnostics (BC and antigen testing), this new multiplex PCR assay may contribute to a rapid and accurate diagnosis of candidiasis.

Background: Vancomycin resistant enterococci (VRE) are an emerging problem in health care settings. The purpose of the investigation was to assess the extent of the outbreak including environmental contamination and to limit further transmission. Methods: We used retrospective patient and laboratory data including pulse field gel electrophoresis (PFGE) typing and virulence and resistance gene analysis. For comparison of medians the Mann-Whitney and for comparison of proportions the Fisher exact tests were used. Results: PFGE typing of VRE strains of an outbreak of 15 VRE cases in a solid transplant unit revealed that nine of the cases belonged to one identical pattern (A), which was only found twice in the environment. Eleven further positive environmental samples showed a different, but identical PFGE pattern E. Only one patient was infected with this environmental strain. Two of nine (22.2%) PFGE A, but nine of eleven (81.2%) PFGE E samples were positive for gelatinase E (p = 0.01), which is described as enhancing biofilm production, suggesting a survival benefit for this strain on inanimate surfaces. Conclusion: Routine disinfection was not able to stop the cluster, but after repeated enforcement of the infection prevention and control (IPC) bundle such as training, strict adherence to hand hygiene and surface disinfection no further cases were observed. We conclude that certain VRE strains predominate in the environment whereas others predominate in humans. Enforcement of the IPC bundle is essential for controlling VRE outbreaks and reducing further transmission.

Wastewater-based epidemiology (WBE) is an effective approach for tracking information on spatial distribution and temporal trends of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) at the community level. Herein, the development, implementation, and operation of the wastewater monitoring program serving Tyrol – a federal province of Austria – are described. The development of this program was initiated by Tyrolean health authorities at the end of the first phase of the Coronavirus disease 2019 (COVID-19) pandemic (May 2020). In close co-operation with the water sector and academic institutions, efficient and effective workflows and processes for wastewater surveillance were established. The monitoring program went into operation in November 2020. By the end of July 2021, a total of 5,270 wastewater influent samples collected at 43 sites were analyzed. The monitoring program provided valuable insights into the development of the pandemic situation in Tyrol and fulfilled several tasks that are of importance in different phases of the pandemic. It represented an early-warning system, provided independent confirmation of temporal trends in COVID-19 prevalence, enabled the assessment of the effectiveness of measures, alerted about bursts of disease activity, and provided evidence for the absence of COVID-19. These findings underline the importance of establishing national wastewater monitoring programs as a complementary source of information for efficient and effective pandemic management.

Blood stream infections pose a major challenge for clinicians as the immediate application of an appropriate antibiotic treatment is the vital factor to safe the patients´ lives. This preliminary study compares three different systems promising fast pathogen identification and susceptibility testing in comparison to conventional blood culture (BC): i) the rapid antimicrobial susceptibility testing protocol according to EUCAST in combination with the Sepsityper^® kit (sRAST), ii) the direct inoculation method on the VITEK^®2 system (dVIT) and iii) testing with the Accelerate Pheno^® system (AccPh). All methods were assessed in terms of accuracy, time to result and usability. Twenty‐three BC samples obtained from patients suffering from proven sepsis were analyzed in detail. Pathogen identification was successful in 95.6%, 91.3% and 91.3% in sRAST, dVIT and AccPh, respectively. Categorical agreement in antimicrobial susceptibility testing was 89.5%, 96% and 96.6%, respectively. Time to result from sample entry to reporting ranged from an average of 4.6 h for sRAST and 6.9 h for AccPh to 10.6 h for dVIT. These results imply a significant shortening of reporting times at considerably high agreement rates for these new diagnostic approaches.

Background: Dermatomycoses, superficial fungal infections of the skin, hair and nails, are among the most common dermatological conditions worldwide. Rapid and accurate diagnosis is essential, particularly in light of emerging antifungal resistance. Conventional diagnostic methods are limited by long turnaround times and lack of species‐level specificity; hence the use of modern DNA‐based tools should be expedient. Objectives: The aim of this study was to evaluate the diagnostic performance of a pan‐dermatophyte PCR‐based workflow for dermatomycosis in routine diagnostics and to describe the epidemiological landscape in Tyrol, Austria. Methods: In this retrospective study, 4483 patient specimens (skin, hair and nails) submitted to the Institute of Hygiene and Medical Microbiology, Medical University of Innsbruck, between 2018 and 2024 were analysed. The workflow included initial pan‐dermatophyte PCR, followed by microscopy and fungal culture for PCR‐negative or inconclusive cases. Species identification was performed by Matrix‐assisted Laser‐Desorption Ionisation Time‐of‐Flight mass spectrometry or, if unsuccessful, by sequencing. Results: Of all specimens, 1170 (26.1%) were PCR‐positive, predominantly with Trichophyton rubrum (76.4%), or members of the T. mentagrophytes‐interdigitale complex (15.4%). In PCR‐negative but microscopy‐positive samples, 67 dermatophytes were identified by culture. The PCR‐based workflow demonstrated a sensitivity of 94.6%, a negative predictive value of 98.0%, and an overall diagnostic accuracy of 98.5%. Among 335 non‐dermatophyte fungi, Aspergillus spp. were most frequent. Conclusion: The proposed workflow demonstrated high sensitivity and accuracy, supporting its suitability for routine diagnostics. It reduced the need for microscopy and culture while enabling reliable species‐level identification, facilitated epidemiological surveillance, revealing a predominance of T. rubrum .

Vancomycin-resistant Enterococcus faecium (VREfm) is an important nosocomial pathogen. The recent emergence of the highly virulent clonal complex 17 (CC17) is posing a challenge for both therapeutic interventions and hospital infection control measures. Hence, prompt discrimination of CC17 VREfm from unrelated and less-virulent VREfm strains is essential for preventing its spread in hospitals and beyond. Between January 2022 and November 2024, 340 VREfm primary isolates have been identified in our lab and underwent genotyping by pulsed-field gel electrophoresis (PFGE) to survey a potential outbreak in the Tyrol region. In addition, whole-genome sequencing (WGS) was performed on a selected subset ( n = 40). To curtail the lengthy time-to-result (TTR) of these methods, a novel typing protocol using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was established, validated, and optimized for rapid sample processing. PFGE and WGS showed that 61.2% of isolates ( n = 208) belonged to a specific VREfm cluster identified as CC17 sequence type (ST) 80 vanA VREfm. A comprehensive MALDI-TOF MS analysis identified a distinct peak pattern specific to this lineage. This phenotypic characterization was used as a novel typing method with excellent performance (sensitivity: 1.00 [0.98−1.00], specificity: 0.89 [0.70−0.97]) and demonstrated a short TTR of 1 day after the cultural growth of VREfm. A rapid and novel MALDI-TOF MS-based typing approach for a specific CC17/ST80 vanA VREfm cluster was developed and enabled real-life application in routine diagnostics to assure accurate infection prevention and control measures. Future outbreak investigations may benefit from adopting this cost- and labor-efficient approach. IMPORTANCE: This study addresses the urgent need for faster ways to detect problematic hospital bacteria. A highly transmissible strain of Enterococcus faecium (CC17) has been spreading in healthcare settings, making infections harder to treat and control. Traditional methods to identify and track outbreaks are accurate but slow and resource-intensive, delaying critical infection control actions. By developing and validating a new method using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, the researchers demonstrated that this strain can be identified quickly, reliably, and at lower cost. Importantly, the new approach delivers results within a day, compared to the lengthy turnaround times of existing methods. This rapid detection tool provides hospitals with a practical solution to respond to outbreaks more effectively, prevent further spread, and protect vulnerable patients. The findings highlight a valuable step forward in strengthening hospital infection control and improving patient safety.