M Bes

Hospices Civils de Lyon, Lyons, Rhône-Alpes, France

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Publications (161)656.9 Total impact

  • Journal of Antimicrobial Chemotherapy 09/2015; DOI:10.1093/jac/dkv278 · 5.31 Impact Factor
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    ABSTRACT: Infective endocarditis (IE)((1)) is a severe condition complicating 10-25% of Staphylococcus aureus bacteremia. Although host-related IE risk factors have been identified, the involvement of bacterial features in IE complication is still unclear. We characterized strictly defined IE and bacteremia isolates and searched for discriminant features. S. aureus isolates causing community-acquired, definite native-valve IE (n=72) and bacteremia (n=54) were collected prospectively as part of a French multicenter cohort. Phenotypic traits previously reported or hypothesized to be involved in staphylococcal IE pathogenesis were tested. In parallel, the genotypic profiles of all isolates, obtained by microarray, were analyzed by discriminant analysis of principal components (DAPC)((2)). No significant difference was observed between IE and bacteremia strains, regarding either phenotypic or genotypic univariate analyses. However, the multivariate statistical tool DAPC, applied on microarray data, segregated IE and bacteremia isolates: IE isolates were correctly reassigned as such in 80.6% of the cases (C-statistic 0.83, P<0.001). The performance of this model was confirmed with an independent French collection IE and bacteremia isolates (78.8% reassignment, C-statistic 0.65, P<0.01). Finally, a simple linear discriminant function based on a subset of 8 genetic markers retained valuable performance both in study collection (86.1%, P<0.001) and in the independent validation collection (81.8%, P<0.01). We here show that community-acquired IE and bacteremia S. aureus isolates are genetically distinct based on subtle combinations of genetic markers. This finding provides the proof of concept that bacterial characteristics may contribute to the occurrence of IE in patients with S. aureus bacteremia. Copyright © 2015. Published by Elsevier B.V.
    Infection, genetics and evolution: journal of molecular epidemiology and evolutionary genetics in infectious diseases 08/2015; DOI:10.1016/j.meegid.2015.08.029 · 3.02 Impact Factor
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    ABSTRACT: Staphylococcus aureus Panton-Valentine leukocidin (PVL) is associated with primary skin and soft-tissue infections (SSTI). We aimed to divert the RIDA®GENE PVL kit (RBiopharm) from its intended use on cultures to the detection of PVL-encoding genes directly from pus samples. Performance was compared with that of the in-house PCR method developed by the French National Reference Centre for Staphylococci. From June 2013 to May 2014, pus samples from S. aureus SSTI were tested. Our in-house PCR was performed on parallel cultures as the gold standard, while the RIDA®GENE PVL assay was used directly on pus samples from the sterile container, or a swab or an Eswab previously dipped in the pus. The kit specificity was also evaluated with pus samples that grew Streptococcus pyogenes. S. aureus reference strains harboring PVL-encoding genes, including known polymorphisms, were also tested. A total of 56 S. aureus-containing pus samples (28 PVL + and 28 PVL-) were collected and analyzed. Sensitivity and specificity of the commercial kit were 96.4 % and 100 % respectively, with equal performance whether tested directly from the sterile container or the Eswab. Sensitivity was lower (67.9 %) when the test was performed from a regular SSTI swab. None of the Streptococcus pyogenes pus samples scored positive (n = 5). Specificity was assessed using reference strains (n = 14); in all strains the PVL gene was correctly detected. This study identified the RIDA®GENE PVL kit as an efficient, sensitive, and specific tool for the rapid detection of PVL-encoding genes in pus samples.
    European Journal of Clinical Microbiology 07/2015; DOI:10.1007/s10096-015-2431-9 · 2.67 Impact Factor
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    ABSTRACT: Staphylococcus aureus bacteremia is associated with high mortality and morbidity, requiring prompt and appropriate antimicrobial treatment. Therefore, it is important to detect methicillin-resistant S. aureus (MRSA) rapidly from blood cultures. Two immunochromatographic tests, BinaxNow S. aureus and BinaxNow PBP2a, were directly applied to 79 Bact/Alert bottles that were positive for Gram positive cocci in cluster aggregations. Sensitivity and specificity for the identification of S. aureus and determination of methicillin resistance were 94% and 87%, and 100% and 100%, respectively, with less than 30 min of performance time. These tests are efficient and rapid; these tests are valuable alternatives to more sophisticated and expensive methods used in the diagnosis of MRSA bacteremia.
    Annals of Laboratory Medicine 07/2015; 35(4):454. DOI:10.3343/alm.2015.35.4.454 · 1.48 Impact Factor
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    ABSTRACT: We describe two cases of human infections caused by Staphylococcus aureus clonal complex (CC) 75, also called Staphylococcus argenteus, harbouring the Panton-Valentine leucocidin (PVL). These two sporadic cases were community-acquired, and identified in France in 2014. Both had an epidemiological link with Mayotte, an overseas department of France located in the Indian Ocean off the south-eastern African coast. This report illustrates that, contrary to previous descriptions, S. argenteus can acquire important virulence factors and be responsible for severe infections.
    Eurosurveillance: bulletin europeen sur les maladies transmissibles = European communicable disease bulletin 06/2015; 20(23). DOI:10.2807/1560-7917.ES2015.20.23.21154 · 5.72 Impact Factor
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    ABSTRACT: Staphylococcus aureus is an opportunistic bacterium capable of causing a wide range of severe diseases when it gains access to underlying tissues. Paradoxically, S. aureus is a common inhabitant of the skin microflora and colonizes the nares and other human mucosa. The purpose of this study was to determine genetic basis for the differences in pathogenic versus colonizing potential of S. aureus isolated from diabetic foot ulcers (DFU). By performing optical map comparisons of a collection of S. aureus strains isolated from DFU, we brought to light a prophage present in non-infecting bacteria. The phage, namely ROSA-like, was localized in a hotspot region ΦNM2 near the locus isd, the iron surface determinant system. The integrated phage reduces significantly the virulence of the strain and increases the biofilm formation. DFU seems to be a specific niche of this colonizing strain. The ROSA-like phage represents the first description of a mobile element present mainly in S. aureus isolated from DFU, which modulates the relationship of the bacteria with its human host. This phage appears to attenuate bacterial virulence and to promote colonization. © 2015 by the American Diabetes Association. Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered.
    Diabetes 04/2015; 64(8). DOI:10.2337/db15-0031 · 8.10 Impact Factor
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    ABSTRACT: Biofilm formation, intra-osteoblastic persistence, small colony variants (SCVs), and the dysregulation of agr, the major virulence regulon, are possibly involved in staphylococcal bone and joint infection (BJI) pathogenesis. We aimed to investigate the contributions of these mechanisms among a collection of 95 Staphylococcus aureus clinical isolates from 64 acute (67.4%) and 31 chronic (32.6%) first episodes of BJI. The included isolates were compared for internalization rate, cell damage, and SCV intracellular emergence using an ex vivo model of human osteoblast infection. Biofilm formation was assessed in a microbead immobilization assay (BioFilm Ring test). Virulence gene profiles were assessed by DNA microarray. Seventeen different clonal complexes were identified among the screened collection. The staphylococcal internalization rate in osteoblasts was significantly higher for chronic than acute BJI isolates, regardless of the genetic background. Conversely, no differences regarding cytotoxicity, SCV emergence, biofilm formation, and virulence gene distribution were observed. Additionally, agr dysfunction, detected by the lack of delta-toxin production using whole cell matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) analysis (n=15; 15.8%), was significantly associated with BJI chronicity, osteoblast invasion, and biofilm formation. These findings provide new insights into MSSA BJI pathogenesis, suggesting the correlation between chronicity and staphylococcal osteoblast invasion. This adaptive mechanism, along with biofilm formation, is associated with agr dysfunction, which can be routinely assessed by delta-toxin detection using MALDI-TOF spectrum analysis, possibly providing clinicians with a diagnostic marker of BJI chronicity at the time of diagnosis. Copyright © 2015. Published by Elsevier Ltd.
    Clinical Microbiology and Infection 02/2015; 21(6). DOI:10.1016/j.cmi.2015.01.026 · 5.77 Impact Factor
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    ABSTRACT: Recent research on Staphylococcus aureus vaccine development has focused on active immunization against Panton–Valentine leukocidin (PVL), a potent leukotoxin associated with both superficial and severe deep-seated infections. PVL prevalence is highly variable worldwide, but it is unknown to what extent immunity to PVL varies between patients from geographic areas with different PVL-positive S. aureus prevalences. We conducted a retrospective multicentric study of anti-PVL and anti-alpha-toxin (Hla) antibody levels in uninfected adult patients from France (low PVL prevalence; n = 200), Algeria (moderate prevalence; n = 143), and Senegal (high prevalence; n = 228). The antibody levels were quantified by an enzyme-linked immunosorbent assay (ELISA) procedure. Because Hla is present in virtually all S. aureus strains, its corresponding antibody levels were considered to reflect population exposure to S. aureus. Compared with French participants, the average anti-PVL antibody levels were 2.5-fold and 8.2-fold higher in Algerian and Senegalese participants, respectively (p S. aureus. Hence, anti-PVL antibody levels in the general populations of France, Algeria, and Senegal vary widely and match variations in PVL-positive S. aureus strain prevalence, with an increasing north-to-south gradient. To conclude, immunity to PVL in a given population correlates with local PVL prevalence. This finding can help to inform PVL vaccine strategies.
    European Journal of Clinical Microbiology 01/2015; 34(5). DOI:10.1007/s10096-014-2307-4 · 2.67 Impact Factor
  • Annual Meeting of the Association-for-Molecular-Pathology (AMP); 11/2014
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    ABSTRACT: Using a large collection of European and North African methicillin-resistant Staphylococcus aureus (MRSA) isolates with a variety of genetic backgrounds and staphylococcal cassette chromosome mec (SCCmec) types, we evaluated the reliability of the BD GeneOhm MRSA assay. Results revealed high performance of this test for detecting MRSA strains provided from Europe and North Africa (98.3%).
    Journal of Clinical Microbiology 10/2014; 52(12). DOI:10.1128/JCM.02139-14 · 3.99 Impact Factor
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    ABSTRACT: SUMMARY Following the recognition of a mecC MRSA isolate from a patient hospitalized in the northeastern region of Slovenia, a national collection of 395 community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) isolates from 2006 to 2013 was screened. An additional six mecC MRSA strains were found and characterized as spa types t843, t9397 and t10009, and multilocus sequence type ST130. The low oxacillin minimum inhibitory concentrations and absence of the mecA gene make recognition of these MRSA strains problematical for diagnostic laboratories. In such strains the presence of mecC should be determined.
    Epidemiology and Infection 07/2014; 143(05):1-4. DOI:10.1017/S0950268814001861 · 2.54 Impact Factor
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    ABSTRACT: Glycopeptides are known to select for heterogeneous vancomycin-intermediate Staphylococcus aureus (h-VISA) from a susceptible strain. In certain clinical situations, h-VISA strains have been isolated from patients without previous exposure to glycopeptides, such as cystic fibrosis patients who frequently receive repeated treatments with beta-lactam antibiotics. Our objective was to determine whether prolonged exposure to beta-lactam antibiotics could induce h-VISA. We exposed 3 clinical vancomycin-susceptible methicillin-resistant Staphylococcus aureus strains to ceftazidime, ceftriaxone, imipenem and to vancomycin (as a control) at sub-inhibitory concentrations for 18 days in vitro. Population analyses showed a progressive increase in vancomycin resistance; seven of the 12 derived strains obtained after induction were classified as h-VISA according to the following criteria: AUC(day 18):AUC(Mu3) ≥ 90% and/or growth on BHI agar with vancomycin 4 mg/L. The derived isolates had a thickened cell-wall proportional to the level of glycopeptide resistance. Genes known to be associated with glycopeptide resistance (vraSR, yvqF, SA1703, graRS, walKR, rpoB) were PCR-sequenced; no de novo mutation was observed upon beta-lactam exposure. To determine whether trfA, a gene encoding a glycopeptide resistance factor, was essential in selecting h-VISA upon beta-lactam pressure, a trfA knock-out strain was generated by allelic replacement. Indeed, beta-lactams exposure of this mutated strain showed no capacity to induce vancomycin resistance. In conclusion, these results showed that beta-lactam antibiotics at subinhibitory concentration can induce intermediate vancomycin resistance in vitro. This induction required an intact trfA locus. Our results suggest that prior use of beta-lactam antibiotics could compromise the vancomycin efficacy for MRSA infections treatment.
    Antimicrobial Agents and Chemotherapy 06/2014; 58(9). DOI:10.1128/AAC.02574-14 · 4.48 Impact Factor
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    ABSTRACT: Objectives Community-acquired Staphylococcus aureus necrotizing pneumonia is a life-threatening disease. Panton Valentine Leukocidin (PVL) has been associated with necrotizing pneumonia. PVL triggers inflammasome activation in human macrophages leading to IL-1β release. IL-1β activates lung epithelial cells to release IL-8. This study aimed to assess the relevance of this inflammatory cascade in vivo and to test the potential of an IL-1 receptor antagonist (IL-1Ra/Kineret) to decrease inflammation-mediated lung injury. Methods We used the sequential instillation of Heat-killed S. aureus and PVL or S. aureus infection to trigger necrotizing pneumonia in rabbits. In these models, we investigated inflammation in the presence or absence of IL-1Ra/Kineret. Results We demonstrated that the presence of PVL was associated with IL-1β and IL-8 release in the lung. During PVL-mediated sterile pneumonia, Kineret/IL-1Ra reduced IL-8 production indicating the relevance of the PVL/IL-1/IL-8 cascade in vivo and the potential of Kineret/IL-1Ra to reduce lung inflammation. However, Kineret/IL-1Ra was ineffective in blocking IL-8 production during infection with S. aureus. Furthermore, treatment with Kineret increased the bacterial burden in the lung. Conclusions Our data demonstrate PVL-dependent inflammasome activation during S.aureus pneumonia, indicate that IL-1 signaling controls bacterial burden in the lung and suggest that therapy aimed at targeting this pathway might be deleterious during pneumonia.
    PLoS ONE 06/2014; 9(6):e97546. DOI:10.1371/journal.pone.0097546 · 3.23 Impact Factor
  • The Journal of Infectious Diseases 05/2014; DOI:10.1093/infdis/jiu246 · 6.00 Impact Factor
  • Journal of Antimicrobial Chemotherapy 04/2014; 69(8). DOI:10.1093/jac/dku130 · 5.31 Impact Factor
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    ABSTRACT: Background. An outbreak of PVL-positive MSSA skin and soft tissue-infections (SSTIs) was suspected in May 2010 when recurrent SSTI was diagnosed in an inmate of a large prison in Nantes, France. Methods and findings. Retrospective and prospective investigations were performed. Microbiological characterisation was by DNA microarray testing (S. aureus genotyping - Identibac, Alere). We identified 14 inmates meeting our clinical and microbiological case definition for PVL-MSSA SSTI between March 2010 and April 2011. The SSTIs developed in tattooed areas in 4 patients and in areas shaved daily with a mechanical razor in 4 other patients. All case isolates exhibited a similar SmaI pulsed-field gel electrophoresis pattern. Microarray analysis showed that all 14 isolates harboured genes encoding PVL and enterotoxins (A, H, K, and Q) and belonged to clonal complex 1 (CC1). Individual and collective hygiene measures, education delivered to inmates and prison employees, and antibiotic treatment of SSTIs were successful in controlling the outbreak. No new cases were identified after April 2011. Routine screening for PVL-positive MSSA carriage was not feasible. Conclusions. Our data suggest that tattooing and shaving with mechanical razors may constitute risk factors for SSTIs among previously colonised inmates and contribute to the PVL-MSSA outbreak in the prison. Allowing inmates access to professional tattooists and to the hygiene and safety conditions available to people in the community would help to prevent tattoo-related infections.
    PLoS Currents 03/2014; 6. DOI:10.1371/currents.outbreaks.e4df88f057fc49e2560a235e0f8f9fea
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    ABSTRACT: The prevalence of CC398 methicillin-susceptible Staphylococcus aureus (MSSA) was unexpectedly high among bone and joint infections (BJIs) and nasal-colonizing isolates in France, with surprising geographical heterogeneity. With none of the major, most-known staphylococcal virulence genes, MSSA CC398 BJI was associated with reduced inflammatory syndrome and lower treatment failure rates. This article is protected by copyright. All rights reserved.
    Clinical Microbiology and Infection 03/2014; 20(10). DOI:10.1111/1469-0691.12567 · 5.77 Impact Factor
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    ABSTRACT: Acquisition of nasal Staphylococcus aureus (S. aureus) colonization by contaminated hands is likely an important determinant of its nasal carriage rate in health care and lab setting. The objective of our cross-sectional study was to assess the prevalence of nasal methicillin-sensitive (MSSA) or -resistant Staphylococcus aureus (MRSA) carriage among health care professionals (HCPs) attending an international symposium and to study the association between compliance with hygiene rules, individual-related parameters, and medical conditions with nasal S. aureus carriage in this population. After obtaining consent, two nasal swabs were collected. Nasal MSSA and MRSA carriage was measured by the: i) molecular approach targeting spa, mecA and mecA-orfX junction sequences, and ii) culture on selective S. aureus media combined with mecA molecular detection of isolated strains. Information on compliance with hygiene rules, demographic variables, sector of activity and long-term medication was collected by anonymous questionnaire. The participation rate was 32.3%. In total, 176 subjects from 34 countries were included in the analysis. S. aureus was isolated from the nasal swabs of 57 (32.4%) subjects, of whom 3 (5.3%) harbored MRSA strains. Overall, 123 subjects reported working in microbiology laboratories with direct manipulation of S. aureus, and 29 acknowledged regular contacts with patients. In this exposed population, hydro-alcoholic solutions appeared to have a significant protective effect against nasal S. aureus carriage (OR = 0.36; 95% CI: 0.15-0.85). Hospital work was associated with increased risk of nasal S. aureus carriage (OR = 2.38; 95% CI: 1.07-5.29). The results of this study showed that compliance with basic rules of hygiene, such as the use of hydro-alcoholic solutions, could reduce the risk of nasal S. aureus colonization. Hydro-alcoholic solution could interrupt auto-transmission of the pathogen, consequently decreasing the overall nasal carriage rate, specifically in transient carriers.
    PLoS ONE 12/2013; 8(12):e82851. DOI:10.1371/journal.pone.0082851 · 3.23 Impact Factor
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    ABSTRACT: Clonal complex 398 livestok-associated-MRSA (CC398 LA-MRSA) clone is described as a major animal pathogen that can also colonize and infect humans. CC398 methicillin susceptible Staphylococcus aureus (CC398 MSSA) is less described. We identified 126 CC398 MSSA strains of human origin within 6380 S. aureus isolates gathered between 2009 and 2011, from the French National Reference Centre for Staphylococci. They were characterized using antimicrobial susceptibility testing, spa typing, DNA microarrays (Identibac S. aureus Genotyping ®, Alere), CC398-specific sequence PCR, ermT (encoding macrolides résistance) PCR. Fifty-three CC398 LA-MRSA collected from French pigs and veal were used as comparators, and phylogenetic relations between human CC398 MSSA and animal CC398 MRSA populations were explored on the basis of spa-typing and DNA microarrays. CC398 MSSA were able to induce a large spectrum of infections (especially skin, bloodstream, and pneumonias). The prevalence rate of this clone was high in MSSA population, i.e., 24.7% in a local prospective study on nasal colonization, and 7.5% in a national prospective study on infective endocarditis. CC398 MSSA isolates were frequently (89%) erythromycin resistant, due to the presence of the ermT gene, a gene not detected in erythromycin resistant CC398 LA-MRSA strains. Expression of staphylococcal complement inhibitor (scn) and the chemotaxis inhibitory protein (chp), was also specific to this population. The CC398 MRSA signature included also a panel of antibiotic resistance genes, especially a type IV or V cassette mec and tetM. CC398 MSSA and CC398 LA-MRSA populations were closely related based on spa-typing and DNA microarrays, with the MRSA strains forming the most derived lineage in phylogenic trees. Both MSSA and MRSA populations may come from common ancestors, which would have evolved in the settings of different selective pressures, explaining the acquisition of ermT, chp and scn for MSSA, and antibiotic resistance genes for MRSA.
    PLoS ONE 11/2013; 8(11):e68462. DOI:10.1371/journal.pone.0068462 · 3.23 Impact Factor
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    ABSTRACT: Background: To characterize the methicillin-resistant Staphylococcus aureus (MRSA) clones present in Istanbul, 102 MRSA isolates collected during a 5-year period at the Istanbul Medical Faculty Hospital were characterized using microarray analysis and phenotypic resistance profiles. Methods: Resistance to methicillin was detected with a cefoxitin disk diffusion assay and confirmed with a MRSA-agar and MRSA detection kit. Antimicrobial susceptibility testing was performed by a disk diffusion assay and interpreted according to the 2012 guidelines of the Antibiogram Committee of the French Society for Microbiology. Decreased susceptibility to glycopeptides was confirmed using the population analysis profile-area under the curve (PAP-AUC) method. The presence of the mecA gene was detected by polymerase chain reaction. Bacterial DNA was extracted according to the manufacturer's recommended protocol using commercial extraction kits. Strains were extensively characterized using the DNA microarray. Results: Isolates were grouped into six clonal complexes. The most frequently detected clone was the Vienna/Hungarian/Brazilian clone (ST239-MRSA-III), which accounted for 53.9% of the isolates. These isolates were resistant to multiple antibiotics, particularly penicillin, tetracycline, rifampicin, kanamycin, tobramycin, gentamicin, levofloxacin, erythromycin, lincomycin and fosfomycin. Furthermore, three isolates were detected by population analysis profile as heterogeneous vancomycin-intermediate S. aureus (hVISA). The UK-EMRSA-15 clone (ST22-MRSA-IV PVL negative) was detected in 9.8% of the isolates and was mainly susceptible to all anti-staphylococcal antibiotics. Seven isolates (6.9%) were positive for PVL genes and were assigned to the CC80-MRSA-IV clone (European CA-MRSA clone, three isolates), ST8-MRSA-IV clone (USA300 clone, two isolates, one ACME-positive) or ST22-MRSA-IV clone (“Regensburg EMRSA” clone, two isolates). All other clones were detected in one to six isolates and corresponded to well-known clones (e.g., Pediatric clone, Dublin EMRSA clone, WA MRSA-54/63, WA MRSA-1/57). Conclusions: This work highlighted both the high prevalence of ST239-MRSA-III clone and the large diversity of the other MRSA clones detected in a university hospital in Istanbul.
    International journal of medical sciences 10/2013; 10(12):1740-5. DOI:10.7150/ijms.6438 · 2.00 Impact Factor

Publication Stats

7k Citations
656.90 Total Impact Points


  • 2007–2015
    • Hospices Civils de Lyon
      Lyons, Rhône-Alpes, France
    • University of Lyon
      Lyons, Rhône-Alpes, France
  • 2014
    • Claude Bernard University Lyon 1
      Villeurbanne, Rhône-Alpes, France
    • Ecole normale supérieure de Lyon
      Lyons, Rhône-Alpes, France
    • French National Centre for Scientific Research
      Lutetia Parisorum, Île-de-France, France
  • 2007–2013
    • French Institute of Health and Medical Research
      Lutetia Parisorum, Île-de-France, France
  • 2011
    • University of Texas Health Science Center at Houston
      • Center for Infectious Diseases
      Houston, Texas, United States
    • Agence Régionale de Santé (ARS)
      Lutetia Parisorum, Île-de-France, France
  • 2007–2011
    • Unité Inserm U1077
      Caen, Lower Normandy, France
  • 2010
    • Statens Serum Institut
      København, Capital Region, Denmark
  • 2006
    • University of Bath
      • Department of Biology and Biochemistry
      Bath, England, United Kingdom
  • 1994
    • Hôpital Louis Pradel
      Lyons, Rhône-Alpes, France
  • 1989–1994
    • Institut Pasteur
      Lutetia Parisorum, Île-de-France, France
  • 1993
    • CHU de Lyon - Groupement Hospitalier Edouard Herriot
      Lyons, Rhône-Alpes, France