Entry of Panton–Valentine leukocidin-positive methicillin-resistant Staphylococcus aureus into the hospital: prevalence and population structure in Heidelberg, Germany 2015–2018

Abstract

Staphylococcus aureus is one of the major pathogens causing community-and healthcare-acquired infections. the presence of the virulence factor panton-Valentine leukocidin (pVL) is associated with recurrent infection and clinical severity and generally regarded as a feature of community associatedmethicillin-resistant Staphylococcus aureus (MRSA). to date, the focus of pVL-positive MRSA in hospitalized patients has been on outbreaks. We aimed to investigate whether pVL-positive MRSA has penetrated the community-hospital barrier by determining the prevalence of pVL in MRSA of hospitalized patients. MRSA strains isolated from patients hospitalized > 48 h in Heidelberg University Hospital between 2015 and 2018 Isolates were analysed for the presence of PVL and subjected to spatyping. PVL-positive MRSA were then characterized by whole genome sequencing. We analysed 740 MRSA isolates in the study period and identified 6.2% (n = 46) PVL-positivity. 32.6% of PVL-positive MRSA met the criteria for nosocomial acquisition. the most frequent clones among the pVL-positive strains were ST80-t044 (21.7%, n = 10/46) and ST8-t008 (19.5%, n = 9/46). WGS identified three possible transmission clusters involving seven patients. in conclusion, we found successful epidemic pVL-positive MRSA clones entering the hospital and causing nosocomial infections. preventive measures and constant surveillance should be maintained to prevent transmissions and clonal outbreaks. Staphylococcus aureus is one of the major causes of community and hospital acquired infections. Methicillinresistance is an ongoing problem locally and globally. Although the overall prevalence of methicillin-resistant S. aureus (MRSA) in Germany is declining 1, MRSA strains harboring the pathogenic marker Panton-Valentine leukocidin (PVL) are isolated more frequently 2. The presence of PVL is relevant, as it is associated with a more severe clinical presentation, often with recurrences, deep and multiple lesions and frequent transmission to contact persons, irrespective of methicillin resistance 3-6. In addition, PVL-positive S. aureus often displays multiple resistances to commonly used antibiotics to treat S. aureus infections 3,7. PVL has been linked to community-associated (CA-) MRSA 6. Although the overall prevalence of PVL in Germany is considered low according to data acquired by Schaumburg et al. 8 , a study by Jappe et al. 9 revealed

Antimicrobial resistance and antibiotic consumption in intensive care units, Switzerland, 2009 to 2018

“…Molecular typing is not routinely performed in Switzerland; this would allow to better discriminate haMRSA from caMRSA. In this context it is important to note that the concept of distinguishing haMRSA and caMRSA has been challenged in recent years, with traditional caMRSA such as USA300 increasingly detected in the healthcare setting and even showing a multi-resistant phenotype [ 33 ]. In contrast to MRSA, GRE have become more frequent, particularly in ICU from eastern Switzerland.…”

Surveillance for Colonization, Transmission, and Infection With Methicillin-Susceptible Staphylococcus aureus in a Neonatal Intensive Care Unit

“…Routine screening samples were processed in the microbiological diagnostics laboratory of the university hospital using the total laboratory automation system (Kiestra; Becton and Dickinson) as published elsewhere, with minor modifications. 15 , 16 Briefly, swabs were inoculated onto a biplate chromogenic medium for S aureus and MRSA detection (CHROMagar Staph aureus/MRSAII; BD Diagnostics) and Columbia blood agar with 5% sheep blood as a growth control for sampling validity. Species identification was performed via matrix-assisted laser desorption ionization (MALDI-TOF; Bruker) using a score of at least 2.0 as a cutoff.…”

“…Genomic DNA was sequenced (MiSeq system; Illumina, Inc; 2 × 250 base pairs [bp] paired-end) as described previously. 16 …”

“…Genomic DNA was sequenced (MiSeq system; Illumina, Inc; 2 × 250 base pairs [bp] paired-end) as described previously. 16 Raw sequences were trimmed for quality using Sickle, version 1.33 (parameters, q > 30; l > 45). 20 The cleaned sequences were then assembled using SPAdes, version 3.13.0.…”

Integrative Analysis of Whole Genome Sequencing and Phenotypic Resistance Toward Prediction of Trimethoprim-Sulfamethoxazole Resistance in Staphylococcus aureus

“…Genomic DNA was extracted from overnight bacterial culture using the DNeasy Blood and Tissue Mini kit (QIAGEN GmbH, Germany). Standard genomic library was prepared from the bacterial DNA and sequenced with the Illumina MiSeq platform (2 × 300 bp paired end), as described elsewhere ( Klein et al, 2020 ). For quality control, raw sequences were trimmed using Sickle 1.33 (parameters, q > 30; 1 > 45).…”