Sainte-Foy-lès-Lyon, France
Sainte-Foy-lès-Lyon, France

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Lemriss H.,University of Lyon | Simoes P.M.,University of Lyon | Lemriss S.,Laboratory of Research and Medical Analysis of the Fraternal of Gendarmerie Royale | Butin M.,Northern Hospital Group | And 4 more authors.
Standards in Genomic Sciences | Year: 2015

Staphylococcus capitis is a coagulase-negative staphylococcus (CoNS) commonly found in the human microflora. Recently, a clonal population of Staphylococcus capitis (denominated NRCS-A) was found to be a major cause of late-onset sepsis (LOS) in several neonatal intensive care units in France. Here, we report the complete genome sequence and annotation of the prototype Staphylococcus capitis NCRS-A strain CR01. The 2,504,472 bp long genome (1 chromosome and no plasmids) exhibits a G+C content of 32.81%, and contains 2,468 protein-coding and 59 tRNA genes and 4 rRNA genes. © 2014 The Author(s).


PubMed | Laboratory of Research and Medical Analysis of the Fraternal of Gendarmerie Royale, Northern Hospital Group, Mohammed V University and University of Lyon
Type: Journal Article | Journal: Standards in genomic sciences | Year: 2014

Staphylococcus capitis is a coagulase-negative staphylococcus (CoNS) commonly found in the human microflora. Recently, a clonal population of Staphylococcus capitis (denominated NRCS-A) was found to be a major cause of late-onset sepsis (LOS) in several neonatal intensive care units in France. Here, we report the complete genome sequence and annotation of the prototype Staphylococcus capitis NCRS-A strain CR01. The 2,504,472 bp long genome (1 chromosome and no plasmids) exhibits a G+C content of 32.81%, and contains 2,468 protein-coding and 59 tRNA genes and 4 rRNA genes.


Rasigade J.-P.,University of Lyon | Rasigade J.-P.,National Reference Center for Staphylococci | Rasigade J.-P.,Northern Hospital Group | Trouillet-Assant S.,University of Lyon | And 19 more authors.
PLoS ONE | Year: 2013

Epidemic community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) is associated with more severe and acute forms of osteomyelitis than healthcare-associated (HA-) MRSA. Although S. aureus is now recognized as a facultative intracellular pathogen, the contribution of osteoblast invasion by CA-MRSA to the pathogenesis of osteomyelitis is unknown. Using an ex vivo model of intracellular infection of human osteoblasts, we demonstrated that CA-MRSA strains of diverse lineages share an enhanced ability to kill infected osteoblasts compared to HA-MRSA. Cytotoxicity comparisons of CA-MRSA isogenic deletion mutants revealed that phenol-soluble modulins (PSMs), a class of membrane-damaging exoproteins that are expressed at higher levels in CA-MRSA than in HA-MRSA, are involved in this osteoblast killing, whereas other major CA-MRSA virulence determinants, the Panton-Valentine leukocidin and alpha-toxin, are not involved. Similarly, functional agr and sarA regulators, which control the expression of PSMs and alpha-toxin, were required for the expression of the intracellular cytotoxic phenotype by CA-MRSA, whereas the saeRS regulator, which controls the expression of alpha-toxin but not PSMs, had no impact on cytotoxicity. Finally, PSM transcript levels determined by quantitative reverse-transcriptase PCR were significantly higher in CA-MRSA than in HA-MRSA strains and associated with cell damage in MRSA-infected osteoblasts. These findings provide new insights into the pathogenesis of severe CA-MRSA osteomyelitis and unravel a novel virulence strategy of CA-MRSA, based on the invasion and subsequent killing of osteoblasts by PSMs acting as intracellular toxins. © 2013 Rasigade et al.


Simoes P.M.,University of Lyon | Simoes P.M.,Northern Hospital Group | Simoes P.M.,National Reference Center for Staphylococci | Rasigade J.-P.,University of Lyon | And 17 more authors.
Antimicrobial Agents and Chemotherapy | Year: 2013

Multiresistant Staphylococcus capitis pulsotype NRCS-A has been reported to be a major pathogen causing nosocomial bacteremia in preterm infants. We report that the NRCS-A strain CR01 harbors a novel 60.9-kb composite staphylococcal cassette chromosome mec (SCCmec) element, composed of an SCCmec with strong homologies to Staphylococcus aureus ST398 SCCmec and of an SCCcad/ars/cop harboring resistance genes for cadmium, arsenic, and copper. Whole-genome-based comparisons of published S. capitis strains suggest that strain CR01 acquired the two elements independently. Copyright © 2013 White.


Trouillet-Assant S.,Northern Hospital Group | Trouillet-Assant S.,University of Lyon | Rasigade J.-P.,Northern Hospital Group | Rasigade J.-P.,University of Lyon | And 8 more authors.
Journal of Clinical Microbiology | Year: 2013

We report that the rates of nasal cocolonization with methicillin- susceptible Staphylococcus aureus and methicillin-resistant coagulase-negative staphylococci can vary widely between patients admitted to different wards within a single hospital. Such cocolonization can greatly influence the performance of molecular methicillin-resistant S. aureus (MRSA) screening tests depending on the methods used and targets selected. © 2013, American Society for Microbiology.


Rasigade J.-P.,University of Lyon | Rasigade J.-P.,Northern Hospital Group | Raulin O.,Northern Hospital Group | Picaud J.-C.,Northern Hospital Group | And 9 more authors.
PLoS ONE | Year: 2012

Background: Coagulase-negative staphylococci, mainly Staphylococcus epidermidis, are the most frequent cause of late-onset sepsis (LOS) in the neonatal intensive care unit (NICU) setting. However, recent reports indicate that methicillin-resistant, vancomycin-heteroresistant Staphylococcus capitis could emerge as a significant pathogen in the NICU. We investigated the prevalence, clonality and vancomycin susceptibility of S. capitis isolated from the blood of NICU infants and compared these data to adult patients. Methodology/Principal Findings: We conducted a retrospective laboratory-based survey of positive blood cultures in NICU infants ≥3 days of age (n = 527) and in adult ICU patients ≥18 years of age (n = 1473) who were hospitalized from 2004 to 2009 in two hospital centers in Lyon, France. S. capitis was the most frequent pathogen in NICU infants, ahead of S. epidermidis (39.1% vs. 23.5% of positive blood cultures, respectively). Conversely, S. capitis was rarely found in adult ICU patients (1.0%) compared to S. epidermidis (15.3%). S. capitis bloodstream isolates were more frequently resistant to methicillin when collected from NICU infants than from adult patients (95.6% vs. 53.3%, respectively). Furthermore, we collected and characterized 53 S. capitis bloodstream isolates from NICU infants and adult patients from six distant cities. All methicillin-resistant S. capitis isolates from NICU infants were clonally related as determined by pulsed-field gel electrophoresis. These isolates harbored a type V-related staphylococcal chromosomal cassette mec element, and constantly showed either vancomycin resistance (37.5%) or heteroresistance (62.5%). Conversely, the isolates that were collected outside of the NICU were genetically diverse and displayed much lower rates of vancomycin resistance and heteroresistance (7.7% and 23.1%, respectively). Conclusions/Significance: A clonal population of methicillin-resistant S. capitis strains has spread into several French NICUs. These isolates exhibit reduced susceptibility to vancomycin, which is the most widely used antimicrobial agent in the NICU setting. © 2012 Rasigade et al.


Gaymard A.,University of Lyon | Gaymard A.,Northern Hospital Group | Pichon M.,University of Lyon | Pichon M.,Northern Hospital Group | And 8 more authors.
International Journal of Antimicrobial Agents | Year: 2016

Methicillin-resistant Staphylococcus aureus (MRSA) is involved in community-acquired and nosocomial diseases. The means of MRSA transmission and dissemination in the community remain uncertain. Studies have shown that public transport systems could be a source of MRSA and may serve as a potential source for community-acquired MRSA infections. This study aimed to investigate MRSA contamination on Lyon's metropolitan network (Métro) in France. Hand-touched surfaces were sampled with sterile swabs (Transystem®) during a 1-day transversal study by collecting 50 samples in seven hub stations and two trains for each of the four Métro lines. Then, during a longitudinal study, one sample was collected twice daily for 30 consecutive days in the busiest and most congested hub station. All swabs were incubated in enrichment medium for 24 h and then each suspension was plated onto a chromogenic selective medium for MRSA. After 24 h at 36 °C, all presumptive MRSA colonies were tested using VITEK® MS to confirm identification as S. aureus as well as by Alere™ PBP2a Culture Colony Test and mecA/mecC PCR to check methicillin resistance. Of the 110 swabs tested, 24 presumptive MRSA colonies were isolated, of which 2 were confirmed as S. aureus by VITEK® MS. These two isolates were tested negative using the PBP2a Culture Colony Test and PCR. Unlike other foreign cities such as Lisbon, the current data suggest a low level of MRSA contamination of hand-touched surfaces on Lyon's Métro. This should be put in perspective with the low level of MRSA colonisation in the French community. © 2016 Elsevier B.V. and International Society of Chemotherapy


Trouillet S.,University of Lyon | Trouillet S.,Northern Hospital Group | Rasigade J.-P.,University of Lyon | Rasigade J.-P.,Northern Hospital Group | And 8 more authors.
Journal of Microbiological Methods | Year: 2011

Flow cytometry is a powerful tool for analyzing the adhesion to and invasion of Staphylococcus aureus (S. aureus) to eukaryotic cells. Established techniques have used bacteria that have been genetically modified to express fluorescent proteins or directly labeled with fluorochromes prior to infection. Such approaches are appropriate in most cases; however, the use of genetically or chemically altered bacteria could introduce a bias when measuring fine differences in adhesion and invasiveness. Here, we describe a combined flow cytometry-based invasion and adhesion assay that does not require the processing of bacteria prior to internalization. This method was performed on osteoblastic MG-63 cells infected with S. aureus reference strain 8325-4 and its invasion-deficient isogenic mutant, which carries deletions in the genes encoding fibronectin-binding proteins A and B. The data from this assay were compared to those obtained using the standard gentamicin protection assay. The results obtained by the two methods were consistent. Moreover, quantification of internalized bacteria was more reproducible using the flow cytometry-based assay than the gentamicin protection assay, which allowed for the simultaneous quantification of host cell adhesion and invasion. © 2011 Elsevier B.V.


PubMed | Eastern Hospital Group, Laboratory of Research and Medical Analysis of the Fraternal of Gendarmerie Royale Rabat, Mohammed V University, Institut Universitaire de France and Northern Hospital Group
Type: | Journal: Frontiers in microbiology | Year: 2016

The multi-resistant


PubMed | Research and Medical Analysis Laboratory of Gendarmerie Royale, Northern Hospital Group, Mohammed V University and University of Lyon
Type: Journal Article | Journal: Genome announcements | Year: 2016

Here, we report the draft genome sequences of methicillin-susceptible Staphylococcus captis pulsotype NCRS-C (CR02 strain) and multiresistant Staphylococcus captis pulsotype NCRS-A (CR07 strain).

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