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Cross-contamination of biological samples during handling and preparation, is a major issue in laboratory setups, leading to false-positives or false-negatives. Sample carryover residue in pipette tips contributes greatly to this issue. Most pipette tips on the market are manufactured with hydrophobic polymers that are able to repel high surface tension liquids, yet they lack in performance when low surface tension liquids and viscous fluids are involved. Moreover, hydrophobicity of pipette tips can result in hydrophobic adsorption of biomolecules, causing inaccuracies and loss in precision during pipetting. Here we propose the use of lubricant-infused surface (LIS) technology to achieve omniphobic properties in pipette tips. Using a versatile and simple design, the inner lumen of commercially available pipette tips was coated with a fluorosilane (FS) layer using chemical vapor deposition (CVD). The presence of FS groups on the tips is confirmed by x-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FTIR) tests. After lubrication of the tips through a fluorinated lubricant, the omniphobicity and repellent behaviour of the tips drastically enhanced which are revealed via static and hysteresis contact angle measurements. The repellency of the lubricant-infused pipette tips against physical adsorption is investigated through pipetting a food coloring dye as well as human blood samples and are compared to the untreated tips. The results show significantly less amount carryover residue when the lubricant-infused tips are utilized compared to commercially available ones. We also demonstrate the lubricant-infused tips reduce bacteria contamination of the inner lumen by 3 to 6-log (over 99%, depending on the tip size) after pipetting up and down the bacteria solution.

Cross-contamination of biological samples during handling and preparation, is a major issue in laboratory setups, leading to false-positives or false-negatives. Sample carryover residue in pipette tips contributes greatly to this issue. Most pipette tips on the market are manufactured with hydrophobic polymers that are able to repel high surface tension liquids, yet they lack in performance when low surface tension liquids and viscous fluids are involved. Moreover, hydrophobicity of pipette tips can result in hydrophobic adsorption of biomolecules, causing inaccuracies and loss in precision during pipetting. Here we propose the use of lubricant-infused surface (LIS) technology to achieve omniphobic properties in pipette tips. Using a versatile and simple design, the inner lumen of commercially available pipette tips was coated with a fluorosilane (FS) layer using chemical vapor deposition (CVD). The presence of FS groups on the tips is confirmed by x-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FTIR) tests. After lubrication of the tips through a fluorinated lubricant, the omniphobicity and repellent behaviour of the tips drastically enhanced which are revealed via static and hysteresis contact angle measurements. The repellency of the lubricant-infused pipette tips against physical adsorption is investigated through pipetting a food coloring dye as well as human blood samples and are compared to the untreated tips. The results show significantly less amount carryover residue when the lubricant-infused tips are utilized compared to commercially available ones. We also demonstrate the lubricant-infused tips reduce bacteria contamination of the inner lumen by 3 to 6-log (depending on the tip size) after pipetting up and down the bacteria solution.

The Omicron SARS-CoV-2 variant of concern is driving the COVID pandemic at a pace never seen before. When the threat of overwhelming healthcare systems balances the hope of achieving herd immunity, providing booster shots and vaccinating fragile individuals can still be attempted in France to prevent death and hospitalizations.

In the context of SARS-Cov-2 virus disease (COVID-19) pandemic, molecular diagnostic tools were rapidly developed as there are fundamental for a rapid detection of infected people. In this context, and in order to optimize the manipulations and reduce the time to get results, we report the successful use of a sampling tool for COVID-19 diagnosis named microFLOQ® Direct (MFD). Hundred upper respiratory specimens sampled from patients with potential COVID-19 were evaluated using MFD, and results were compared to the results obtained by standard sampling procedure using dry swabs and physiologic serum as the transport medium. MFD results compared to results issued from the classic RNA purification and amplification steps from transport medium showed that MFD can be directly used for RT-PCR analysis without the preliminary inactivation and extraction steps. So, MFD could limit handling errors compared to the different treatment steps with dry swabs and transport medium. It therefore limits the risk of contamination, simplify the analytical process and enables to get results in less than 2 hours. We also show that the MFD kit is operational as a screening tool in the field of molecular detection of viral and bacterial diseases during an outbreak and then it can be used for public health or agro-veterinary purposes.

Cross-contamination of biological samples during handling and preparation, is a major issue in laboratory setups, leading to false-positives or false-negatives. Sample carryover residue in pipette tips contributes greatly to this issue. Most pipette tips on the market are manufactured with hydrophobic polymers that are able to repel high surface tension liquids, yet they lack in performance when low surface tension liquids are involved. This presents an obstacle for pipette tips as inaccuracies and loss in precision arise when low surface tension liquids such as viscous oils are pipetted. Here we propose the use of lubricant-infused surface (LIS) technology to achieve omniphobic properties in pipette tips. Using a versatile and simple design, the inner lumen of commercially available pipette tips was coated with a fluorosilane (FS) layer using chemical vapor deposition (CVD). We show that after lubricating the tips through simply pipetting up and down a fluorinated lubricant, the surface free energy of the tips drastically decreased enabling them to attain low retention properties. Contact angle measurements reveals that the treated pipette tips have enhanced omniphobic properties. The repellent behavior of the lubricant-infused pipette tips against physical adsorption is investigated through pipetting a food coloring dye as well as human blood samples and are compared to the untreated tips. The results show significantly less amount carryover residue when the lubricant-infused tips are utilized compared to commercially available ones.

Blood culture bottles (BCBs) are commonly used for the diagnosis of infections associated with orthopedic devices. Although Cutibacterium acnes is an important pathogen in orthopedics, relatively little is known about its growth characteristics in BCBs. This prompted us to analyze the influence of bacterial genotype and clinical significance on time-to-detection (TTD) in BCBs.We reviewed 59 cases of orthopedic device-related infections in which at least one intraoperative specimen yielded a pure C. acnes culture from anaerobic BCBs (BD Bactec Lytic/10 Anaerobic/F; Lytic-Ana) and/or solid media. A strain was considered infectant if the same genotype was present in two or more intraoperative samples. From these cases, we isolated a total of 72 unique C. acnes strains belonging to four multilocus sequence type clonal complexes (CCs): CC18, CC28, CC36 and CC53. Growth rate and TTD in Lytic-Ana BCB were studied under experimental conditions (inoculation of standard inoculum) and in clinical samples (inoculation of periprosthetic tissue samples).Median TTD values were shorter for CC53 compared to other CCs under experimental conditions (69 vs. 103 h; p 0.001) and from clinical specimens (70 vs. 200 h; p = 0.02). Infectant strains had a shorter median TTD compared to contaminant strains in a clinical situation, while the difference was not observed under experimental conditions.The detection dynamics of C. acnes in Lytic-Ana BCBs were associated with genotype. Thus, TTD not only reflects the bacterial load in clinical samples, but may also reflect the intrinsic properties of the clonal complex of C. acnes.

The increasing incidence of carbapenemase-producing Gram-negative bacilli (C-PGNB) represents a major public health challenge. Rapid detection of digestive colonization with C-PGNB is fundamental to control their spread. We performed the validation of a rapid protocol for C-PGNB detection directly on rectal swabs. We developed a protocol combining enrichment by a rapid selective subculture of the rectal swab medium and realization of a Resist-4 O.K.N.V. K-SeT test on the bacterial pellet obtained. The limit of detection and performances of this protocol were validated in vitro on 52 C-PGNB strains spiked on a calibrated sample suspension and confirmed in clinical settings on 144 rectal swabs sampled from patients with C-PGNB digestive colonization (n = 48) and controls (patients with extended-spectrum beta-lactamase [ESBL] colonization [n = 48] and without carbapenemase/ESBL [n = 48]). The protocol detected, with 100% sensitivity, the presence of the 15 OXA-48-, 14 KPC-, 13 NDM-, and 10 VIM-producing GNB from 10(3) CFU/ml. The limit of detection was 2 × 10(2) CFU/ml. Among the 48 C-PGNB-containing rectal swabs of the validation cohort, 46 were accurately detected. False negative were observed for 1 NDM-producing Acinetobacter baumannii strain and 1 OXA-48-producing Escherichia coli strain. The 96 control swabs were negative. Sensitivity and specificity for C-PGNB detection were 97.7% (95% confidence interval [CI], 87.7 to 100) and 100% (95% CI, 96.2 to 100). The negative likelihood ratio was 0.04 (95% CI, 0.01 to 0.16). Considering a C-PGNB digestive colonization prevalence between 0.01% and 0.1%, positive and negative predictive values were 100%. Our protocol is a rapid and low-cost method detecting accurately the digestive colonization with carbapenemase-producing Enterobacteriaceae in 4 h without any requirement for specific equipment.

The emergence and quick spread of SARS-CoV-2 has pointed at a low capacity response for testing large populations in many countries, in line of material, technical and staff limitations. The traditional RT-qPCR diagnostic test remains the reference method and is by far the most widely used test. These assays are limited to a few probe sets, require large sample PCR reaction volumes, along with an expensive and time-consuming RNA extraction step. Here we describe a quantitative nanofluidic assay that overcomes some of these shortcomings, based on the BiomarkTM instrument from Fluidigm. This system offers the possibility of performing 4608 qPCR end-points in a single run, equivalent to 192 clinical samples combined with 12 pairs of primers/probe sets in duplicate, thus allowing the monitoring of SARS-CoV-2 including the detection of specific SARS-CoV-2 variants, as well as the detection other pathogens and/or host cellular responses (virus receptors, response markers, microRNAs). The 10 nL-range volume of BiomarkTM reactions is compatible with sensitive and reproducible reactions that can be easily and cost-effectively adapted to various RT-qPCR configurations and sets of primers/probe. Finally, we also evaluated the use of inactivating lysis buffers composed of various detergents in the presence or absence of proteinase K to assess the compatibility of these buffers with a direct reverse transcription enzymatic step and we propose several protocols, bypassing the need for RNA purification. We advocate that the combined utilization of an optimized processing buffer and a high-throughput real-time PCR device would contribute to improve the turn-around-time to deliver the test results to patients and increase the SARS-CoV-2 testing capacities.

Immune checkpoint inhibitors (ICIs) are commonly used in patients with advanced non-small cell lung cancer (NSCLC). An unmet need remains for new biomarkers associated with ICIs. In this study, consecutive patients with advanced NSCLC treated with nivolumab or pembrolizumab were included. Plasma at ICIs initiation was prospectively collected and a multiplex ELISA assay testing 48 cytokines and growth factors was performed. Exploratory endpoints were the association between plasma biomarkers with outcome and grade III&ndash, IV immune related adverse events (irAEs). Thirty-five patients were included. Patients without clinical benefit (n = 22) had higher pre-ICI soluble Hepatocyte Growth Factor (sHGF) (210.9 vs. 155.8 pg/mL, p = 0.010), lower pre-ICI soluble Fibroblast Growth Factor (sFGF) (4.0 vs. 4.8 pg/mL, p = 0.043) and lower pre-ICI interleukine-12 (IL-12) (1.3 vs. 2.2 pg/mL, p = 0.043) concentrations. Patients with early progression (n = 23) had higher pre-ICIs sHGF (206.2 vs. 155.8 pg/mL, p = 0.025) concentrations. Patients with low sHGF levels at ICIs initiation had longer progression-free survival and overall survival than those with high sHGF levels: respectively 2.5 vs. 8.0 months (p = 0.002), and 5.5 vs. 35.0 months (p = 0.001). TNF-&alpha, IL-16, IL-12p40 and MCP3 were associated with high grade irAEs. This study shows the potential association between several plasma biomarkers with outcome and grade 3&ndash, 4 IrAEs in advanced NSCLC treated with ICIs.

International audience; BACKGROUND: Preoperative synovial fluid culture is pivotal in the early diagnosis of prosthetic joint infection (PJI) but may yield false-positive and false-negative results. We evaluated the predictive value of synovial fluid culture results combined with the measurement of serum anti-staphylococcal antibodies (SASA). QUESTIONS/PURPOSES: (1) For hip and knee PJI, does combining positive SASA results with preoperative synovial culture results improve the positive predictive value (PPV) of preoperative synovial fluid culture alone? (2) Does combining preoperative synovial fluid culture results with a positive cell count and differential result increase the PPV of preoperative synovial fluid culture alone? (3) What proportion of isolated organisms exhibit concordance in antibiotic susceptibility: preoperative aspiration versus intraoperative isolates? METHODS: A prospective study was conducted at two French reference centers that manage bone and joint infections and included 481 adult patients who had a revision or resection arthroplasty between June 25, 2012 and June 23, 2014. Exclusion criteria including no serum sample available for immunoassay, the lack of microbiological documentation, and the absence of preoperative aspiration reduced the patient number to 353. Seven patients with an undetermined SASA result were excluded from the analysis. We also excluded patients with PJI involving more than one Staphylococcus species (polystaphylococcal infection) and those in whom more than one Staphylococcus species was recovered from the preoperative synovial fluid culture (polystaphylococcal synovial fluid culture). In total, 340 patients were included in the analysis (no infection, 67% [226 of 340]; staphylococcal infection, 21% [71 of 340]; other infection, 13% [43 of 340]). The preoperative synovial fluid analysis included a cell count and differential and bacterial culture. SASAs were measured using a multiplex immunoassay. The diagnosis of PJI was determined using the Infectious Diseases Society of America (IDSA) criteria [] and intraoperative tissue culture at the time of revision surgery was used as the gold standard (at least one positive intraoperative sample for a "virulent" organism (such as S. aureus) or two positive samples for a "non-virulent" (for example S. epidermidis). RESULTS: SASA increased the PPV compared with synovial fluid culture alone (92% [95% CI 82 to 97] versus 79% [95% CI 68 to 87]; p = 0.04); when stratified by site, an increase in PPV was seen in hip infections (100% [95% CI 89 to 100] versus 77% [95% CI 63 to 88]; p = 0.01) but not in knee infections (84% [95% CI 66 to 95] versus 80% [95% CI 64 to 91]; p = 0.75). A positive cell count and differential result increased the PPV of staphylococcal synovial fluid cultures compared with synovial fluid culture alone (86% [95% CI 70 to 95] versus 79% [95% CI 68 to 87]; p = 0.36); when stratified by site, no difference in hip and knee infections was observed (86% [95% CI 67 to 96] versus 77% [95% CI 63 to 88]; p = 0.42) and 86% [95% CI 70 to 95] versus 80% [95% CI 64 to 91]; p = 0.74). CONCLUSION: SASA measurement improves the predictive value of synovial fluid cultures of the hip for all staphylococcal organisms, including coagulase-negative staphylococci, but the PPV of SASA plus synovial fluid culture it is not superior to the PPV of synovial fluid cell count/differential plus synovial culture for the knee. LEVEL OF EVIDENCE: Level III, diagnostic stud

Corynebacterium striatum is a ubiquitous colonizer of human skin and mucous membranes. It is increasingly involved in infections, especially with prosthetic devices or in immunocompromised individuals. Microbiological diagnosis is challenging and bacterial resistance is a major concern. We performed a retrospective study of monomicrobial bone and joint infections (BJI) due to C. striatum in two referral centers from April 2012 to July 2017. We collected the patients' clinical and microbiological characteristics and outcomes. We also performed a literature review of BJI due to C. striatum. We identified 12 cases (nine prosthetic joint infections, one osteosynthetic device infection, one non-union, and one arthritis) in 11 patients, five of which were immunocompromised. Microbiological diagnosis was performed with prolonged culture media. Ten out of 12 strains were susceptible to aminopenicillin, a drug class not recommended for testing by the EUCAST/CASFM guidelines, and 8/12 patients were treated with amoxicillin-rifampicin. The cure rate was 8/12, after a median follow-up period of 487.5 days (IQR 140.3-1348.5). Twelve cases of BJI due to C. striatum were previously reported. Among them, 5/12 patients were immunocompromised, 3/12 cases were acute BJI, and 2/12 were device-related infections. The diagnosis was performed by PCR in one case, and 10/12 patients were treated with glycolipopeptides, with a cure rate of 11/12. We report the largest cohort of monomicrobial BJI with C. striatum. Determination of aminopenicillin susceptibility is essential since it is frequently active in our experience, even in BJI. The cure rate of this infection seems high.

Cutibacterium acnes is a major etiologic agent of orthopaedic implant-associated infections (IAIs) and requires up to 14 days of incubation in an anaerobic atmosphere for growth detection. As blood culture (BC) systems are increasingly being used to monitor the growth of IAI specimens, we compared different BC media for growth detection of C. acnes. Non-duplicate C. acnes isolates (n = 99) obtained from sonicate-fluid cultures of orthopaedic IAIs from Slovenia (n = 54), conventional tissue samples of monomicrobial orthopaedic IAIs from France (n = 43) and two reference strains were inoculated to anaerobic BC bottles of two major BC systems and 3 conventional culture media types (thioglycolate broth, Schaedler and chocolate agar). Growth and time-to-detection (TTD) were recorded. Only Lytic (BACTEC) and SN (BacT/ALERT) bottles consistently detected growth of C. acnes within 14 days with 94% (n = 93) and 92% (n = 91) detection rates, respectively (p = 0.79). Lytic was superior to Plus BACTEC medium (p < 0.001), while SN was superior to all other BacT/ALERT media (p < 0.001). Mean TTD was 128 ± 43 h (61–336 h) for Lytic and 158 ± 65 h (77–336 h) for SN medium. Among the conventional media, 99% (n = 98) of the isolates grew on Schaedler agar, 96% (n = 95) in thioglycolate broth and 74% (n = 73) on chocolate agar. Inconsistent growth of C. acnes in different BC media can critically influence the detection of this major IAI pathogen. Only Lytic (BACTEC) and SN (BacT/ALERT) BC media types were consistently able to detect C. acnes within 14 days of incubation. However, visible growth was observed faster in thioglycolate broth and Schaedler agar media.

Introduction: Multiplex-antibody detection has been recently proposed for the noninvasive diagnosis of staphylococcal prosthetic joint infection (PJI). We evaluated this approach for the post-treatment follow-up of patients. Methods: Nineteen cases of staphylococcal PJI were prospectively followed for one year after treatment. The IgG response against eight staphylococcal antigens was measured before surgery and one year post-surgery using Luminex technology (Austin, TX, USA); median fluorescence intensity values determined for each antigen were transformed into a “Total Response Index” (TRI). Results: Patients (11 women/8 men) had a mean (SD) age of 72.2 (12.4) years. Site of prosthesis was the knee (n=10), the hip (n=8) and the shoulder (n=1). Ten patients were infected by S. epidermidis, six by S. aureus, and three by S. lugdunensis. TRI values at one year were significantly lower than pre-surgery values (mean [SD]: 5.9 [1.8] versus 8.1 [3.4], p=0.02) and decreased, on average, by 21.2%. TRI values markedly increased in two patients. One patient had a relapse of S. aureus PJI at five months post-surgery, with a 37% increase of the TRI. The other had septic failure three months after revision for S. lugdunensis PJI; all intraoperative samples remained culture-negative, but the TRI increased by 51% and the antibody profile showed a marked change, suggesting a reinfection with another staphylococcal species. Conclusion: Multiplex-antibody measurement may be useful for the follow-up of staphylococcal PJI and may help to detect septic failure involving organisms targeted by the assay.

We developed and evaluated a multiplex antibody detection-based immunoassay for the diagnosis of prosthetic joint infections (PJIs). Sixteen protein antigens from three Staphylococcus species ( Staphylococcus aureus , Staphylococcus epidermidis , and Staphylococcus lugdunensis ) (8 antigens), Streptococcus agalactiae (4 antigens), and Propionibacterium acnes (4 antigens) were selected by comparative immunoproteomics using serum samples from PJI cases versus controls. A bead-based multiplex immunoassay that measured serum IgG against purified, recombinant forms of each of the 16 antigens was developed. We conducted a prospective study to evaluate the performance of the assay. A PJI was defined by the presence of a sinus tract and/or positive intraoperative sample cultures (at least one sample yielding a virulent organism or at least two samples yielding the same organism). A total of 455 consecutive patients undergoing revision or resection arthroplasty (hip, 66.3%; knee, 29.7%; shoulder, 4%) at two French reference centers for the management of PJI were included: 176 patients (38.7%) were infected and 279 (61.3%) were not. About 60% of the infections involved at least one of the species targeted by the assay. The sensitivity/specificity values were 72.3%/80.7% for targeted staphylococci, 75%/92.6% for S. agalactiae , and 38.5%/84.8% for P. acnes . The assay was more sensitive for infections occurring >3 months after arthroplasty and for patients with an elevated C-reactive protein (CRP) or erythrocyte sedimentation rate (ESR). However, it detected 64.3% and 58.3% of targeted staphylococcal infections associated with normal CRP and ESR values, respectively. This new multiplex immunoassay approach is a novel noninvasive tool to evaluate patients suspected of having PJIs and provides information complementary to that from inflammatory marker values.

Background Complement pathway inhibition may provide benefit for severe acute respiratory illnesses caused by viral infections such as COVID-19. We present results from a nonrandomized proof-of-concept study of complement C5 inhibitor eculizumab for treatment of severe COVID-19. Methods All patients (N = 80) with confirmed SARS-CoV-2 infection and severe COVID-19 admitted to our intensive care unit between March 10 and May 5, 2020 were included. Forty-five patients were treated with standard care and 35 with standard care plus eculizumab through expanded-access emergency treatment. The prespecified primary outcome was day-15 survival. Clinical laboratory values and biomarkers, complement levels, and treatment-emergent serious adverse events (TESAEs) were also assessed. Findings At day 15, estimated survival was 82.9% (95% CI: 70.4%‒95.3%) with eculizumab and 62.2% (48.1%‒76.4%) without eculizumab (log-rank test, P = 0.04). Patients treated with eculizumab experienced a significantly more rapid decrease in lactate, blood urea nitrogen, total and conjugated bilirubin levels and a significantly more rapid increase in platelet count, prothrombin time, and in the ratio of arterial oxygen tension over fraction of inspired oxygen versus patients treated without eculizumab. Eculizumab-associated changes in complement levels, laboratory values, and biomarkers were consistent with terminal complement inhibition, reduced hypoxia, and decreased inflammation. TESAEs of special interest occurring in >5% of patients treated with/without eculizumab were ventilator-associated pneumonia (51%/24%), bacteremia (11%/2%), gastroduodenal hemorrhage (14%/16%), and hemolysis (3%/18%). Interpretation Findings from this proof-of-concept study suggest eculizumab may improve survival and reduce hypoxia in patients with severe COVID-19. Randomized studies evaluating the efficacy and safety of this treatment approach are needed. Funding Programme d'Investissements d'Avenir: ANR-18-RHUS60004.

Cutibacterium acnes orthopedic device-related infections (ODRIs) range from obvious infections to solely culture-based diagnoses. Multilocus sequence typing of multiple isolates from the same procedure revealed that most cases with normal C-reactive protein levels that were classified as C. acnes ODRI would be considered contaminations when accounting for genotypic data.

Introduction Deux nouveaux antibiotiques associant une cephalosporine et un inhibiteur de betalactamase ont ete recemment mis sur le marche. Leur efficacite au cours des infections osteo-articulaires est mal connue. Materiels et methodes Etude retrospective monocentrique, de 05/2016 a 01/2019, des prescriptions de ceftolozane-tazobactam (C/T) et ceftazidime-avibactam (C/A) au cours des infections osteo-articulaire. Les patients ont ete identifies par la pharmacie et les dossiers medicaux revus avec une fiche standardisee de recueil incluant indication, caracteristiques cliniques, biologiques et microbiologiques, evolution, tolerance. Resultats Nous avons recense 10 prescriptions de C/T et 14 de C/A. L’âge median etait de 63,0 ans dans le groupe C/T et 64,0 ans (IQR 51,75–66,0) dans le groupe C/A (IQR 44,8–71,0), avec un sexe ratio (M/F) exclusivement masculin dans le groupe C/T et de 2,5 dans le groupe C/A. La duree d’hospitalisation etait de 48,0 jours dans le groupe C/T et de 95,5 jours dans le groupe C/A. Dans le groupe C/A, 13 patients ont beneficie d’une prise en charge medico-chirurgicale pour infection de prothese (3), arthrite (8), escarre avec osteite (3). Dans le groupe C/T, 9 patients ont beneficie d’une prise en charge medico-chirurgicale pour infection de materiel (2), arthrite (5), escarre (2), osteite (1). Les prelevements per-operatoires ont permis un diagnostic microbiologique dans 90 % des cas dans le groupe C/T et dans 92,9 % des cas dans le groupe C/A. Les prelevements etaient polymicrobiens dans 66 % des cas dans le groupe C/T et 85,7 % dans le groupe C/A. Le choix de l’antibiotherapie etait lie a la resistance microbiologique dans 80 % des cas dans le groupe C/T et dans 78,5 % des cas dans le groupe C/A. Dans le groupe C/A, les principales bacteries multiresistantes identifiees etaient : P.aeruginosa (5), K. pneumoniae BLSE (5), K.pneumoniae secretrice de carbapenemase (2), E.coli BLSE (2), E.cloacae BLSE (3), et A. baumanii (1). Dans le groupe C/T, il s’agissait de P. aeruginosa (7), K. pneumoniae BLSE (1), M.morganii (1), C. koseri (1). Dans le groupe C/A, l’evolution etait favorable pour 9 patients, 4 ont rechute et 1 est decede. Dans le groupe C/T, on note 7 guerisons, 2 rechutes et 1 deces. Une encephalopathie sous C/A a ete recensee. Conclusion Peu de donnees sont disponibles sur l’utilisation hors AMM de C/A et C/T dans les infections-osteo-articulaires. Notre experience recense des infections polymicrobiennes avec au moins un germe multi-resistant, principalement P.aeruginosa ou enterobacteries BLSE. Bien que s’agissant de patients multi-operes aux nombreuses comorbidites, l’utilisation de C/A et C/T a permis une reponse favorable lors de situations complexes.

We compared two walk-away molecular diagnostic assays, the GeneXpert MRSA Gen 3 assay and the BD-Max MRSA XT assay. A total of 119 prospective swabs and 36 culture-positive samples were tested. Xpert MRSA Gen 3 had sensitivity of 95.7% and specificity of 100% versus 87.5% and 97.1% for BD-Max. The difference in agreement with the enriched culture results was significantly in favor of the Xpert assay ( P < 0.02, McNemar nonparametric text).

Mycobacterium abscessus is a pathogenic, rapidly growing mycobacterium involved in pulmonary and cutaneo-mucous infections worldwide, to which cystic fibrosis patients are exquisitely susceptible. The analysis of the genome sequence of M. abscessus showed that this bacterium is endowed with the metabolic pathways typically found in environmental microorganisms that come into contact with soil, plants, and aquatic environments, where free-living amoebae are frequently present. M. abscessus also contains several genes that are characteristically found only in pathogenic bacteria. One of them is MAB_0555 , encoding a putative phospholipase C (PLC) that is absent from most other rapidly growing mycobacteria, including Mycobacterium chelonae and Mycobacterium smegmatis . Here, we report that purified recombinant M. abscessus PLC is highly cytotoxic to mouse macrophages, presumably due to hydrolysis of membrane phospholipids. We further showed by constructing and using an M. abscessus PLC knockout mutant that loss of PLC activity is deleterious to M. abscessus intracellular survival in amoebae. The importance of PLC is further supported by the fact that M. abscessus PLC was found to be expressed only in amoebae. Aerosol challenge of mice with M. abscessus strains that were precultured in amoebae enhanced M. abscessus lung infectivity relative to M. abscessus grown in broth culture. Our study underlines the importance of PLC for the virulence of M. abscessus . Despite the difficulties of isolating M. abscessus from environmental sources, our findings suggest that M. abscessus has evolved in close contact with environmental protozoa, which supports the argument that amoebae may contribute to the virulence of opportunistic mycobacteria.

Scedosporium apiospermum is a ubiquitous filamentous fungus, commonly found in soil, sewage and polluted waters. It is rarely pathogenic but can cause a broad spectrum of clinical diseases, which can be localised or disseminate to distant organs. The disseminated form of the disease is mostly seen among immunocompromised patients. However, some rare cases of disseminated disease have been reported in immunocompetent individuals. Treatment of these infections is challenging because of their natural resistance to many antifungal agents. Here, we report the case of a 57-year-old immunocompetent patient diagnosed with femoral pseudarthrosis due to S. apiospermum, despite having no obvious clinical sign of infection. Previously, the patient had undergone four iterative femoral surgeries following a road traffic accident which occurred 20 years before. During its last surgery for pseudarthrosis, no clinical or biological signs of infection were present. Per operative samples tested positive for S. apiospermum. The patient was successfully treated with oral voriconazole during 6 months with an excellent tolerance. We also provide a review of literature on bone and joint infections due to Scedosporium spp. (S. apiospermum, Scedosporium boydii and Scedosporium aurantiacum), discussing the evolution of their management and outcome which seems to improve since the use of voriconazole.

Background The bacterial colonization of urinary catheters is a major source of hospital acquired urinary tract infections (HAUTI). Bacteria repellent coatings could lower HAUTI prevalence and minimize antimicrobial usage. We report a model of spontaneous bacterial colonization of an intravesical catheter in a spinalized rat model, and the first in vivo proof of the efficacy of lubricant-infused catheters (LICs) in preventing bacterial colonization. Methods LICs preparation: Oxygen plasma treated polyethylene catheters were immediately placed in a vacuum desiccator and 200 µL of trichloro (1 hour,1H,2H,2H-perfluorooctyl) silane) was placed beside the catheter segments. The vacuum pump connected to the desiccator was turned on with the exit valve closed once a pressure of −0.08 MPa was achieved. The chemical vapour deposition process was initiated for 4 hours. Catheters were removed from the desiccator and placed in an oven at 60oC for a minimum of 12 hours in order to complete the modification process. Catheters were saturated with a biocompatible fluorocarbon-based lubricant (perfluorodecalin) prior to implantation. Thirty centimeters long native catheters and LICs were surgically implanted in the bladder of rats spinalized 19 days prior and programmed to undergo cystometry experiments 48 hours later. Each rat was maintained individually in a cage with food and water ad libitum until bladder functional evaluation, and benefitted from a of trimethoprim sulfadoxine and fluoroquinolone prophylaxis. At the end of the cystometry experiments, the animals were euthanized and the bladder catheter was removed. A 1 cm section from the intravesical end of the catheter was cut and placed in 1 mL of Amies medium (eSawb, Copan), vortexed and sonicated. Ten microliters of the suspension were plated on URIselect4 medium (Bio-Rad, Hercules, Ca) for bacterial enumeration. Results A significant reduction in bacterial colonization with Enterobacteriaceae and Enterococci was observed in the LICs group (N = 6, below 100 CFU threshold) compared with the native catheter group (N = 5, average 8200 CFU) (P < 0.02). Conclusion Lubricant-infused catheters effectively prevent bacterial colonization in vivo and provide an attractive and nonselective option for HAUTI prevention. Disclosures All authors: No reported disclosures.

Introduction Au cours de la prise en charge en 2 temps des infections sur prothese osteoarticulaires (IPOA), lors de la repose, une antibiotherapie postoperatoire large spectre probabiliste est systematiquement prescrite en raison du possible risque de sepsis persistant. Elle est debutee apres realisation des prelevements a visee microbiologiques peroperatoires et arretee en cas de negativite ; ce qui est frequemment le cas. Un test de detection rapide (15 minutes) de l’α-defensine permettant de detecter les IPOA a ete recemment commercialise et ses resultats semblent prometteurs en cas de suspicion d’infection. Nous nous sommes interesses a sa performance lors des reposes au cours de prise en charge en 2 temps. Il pourrait alors permettre d’eviter une exposition antibiotique large spectre inutile. Materiels et methodes Etude prospective monocentrique de 16 cas de repose de prothese osteoarticulaires post-episode septique ne presentant pas de signe clinique ni biologique d’infection lors du geste. Un test de detection de l’α-defensine a ete realise en extemporane sur le liquide articulaire en peroperatoire avant administration d’antibiotique et en meme temps que les prelevements a visee microbiologique. Nous avons compare les resultats du test d’α-defensine et les cultures microbiologiques prolongees en milieu enrichi apres broyage comme realise dans notre centre. Resultats Au total, 16 patients ont ete inclus, l’âge moyen etait de 67,1 ans (range 43–80), le sex-ratio etait de 1,00. Deux patients etaient immunodeprimes. Il s’agissait de 9 cas de protheses de genou, 4 de hanche, 2 de cheville, et 1 d’epaule. Le nombre moyen d’interventions prealables etait de 2,4. Les bacteries identifiees lors du precedent episode septique etaient : Escherichia coli (n = 2), SAMS (n = 2), SARM (n = 1), staphylocoques a coagulase negative (n = 7) (Staphylococcus warneri, S. lugdunensis, S. haemolyticus, S. simulans, S. schleferi, S. epidermidis meti-R et S. caprae), streptocoques (n = 2), enterocoques (n = 2), anaerobes (n = 1), et Candida sp. (n = 1). Au moment de la reprise les patients etaient apyretiques, il n’existait pas de signe locaux et/ou generaux de sepsis. La CRP moyenne etait de 39,7 mg/L et le taux moyen de leucocytes de 9,6 G/L. Au total, l’α-defensine etait negative dans tous les cas, ainsi que les prelevements microbiologiques. Conclusion L’α-defensine semble un outil interessant pour dedouaner la persistance d’une infection dans le cas de repose de prothese osteoarticulaire et permettrait d’eviter d’exposer inutilement les patients a une antibiotherapie intempestive. Cette indication originale doit etre validee par des etudes de plus large ampleur.

Introduction Cotrimoxazole (Sulfamethoxazole-Trimethoprim, SXT) has interesting characteristics for the treatment of bone and joint infection (BJI): a broad spectrum of activity with adequate bone diffusion and oral and intravenous formulations. However, its efficacy and safety in BJIs are poorly documented and its use remains limited. Methods We conducted a retrospective study in 2 reference centers for BJIs from 2013 to 2018 among patients treated with SXT for a BJI. Data were collected from patient's medical charts. Outcomes and adverse events were evaluated at day (D)7, D45 and D90. Results We analyzed 51 patients with a mean age of 60 ± 20 (SD) years of which 76% presented with an orthopedic device infection (ODI). Gram-negative bacilli (GNB) were involved in 47% of BJIs (n = 24). Moreover, they were often polymicrobial infections (41%). Doses of SXT ranged from 800/160mg bid (61%; n = 31) to 800/160mg tid (39%; n = 20). Median SXT treatment duration was 45 days (IQR 40-45). SXT was part of a dual therapy in 84% of patients (n = 43), associated mainly with fluoroquinolones (n = 17) or rifampicin (n = 14). Outcome was favorable at D7 in 98% (n = 50), at D45 in 88.2% (n = 45) and at D90 in 78.4% (n = 40). The second agent combined with SXT was not an independent factor of favorable outcome (p = 0.97). Adverse events were reported in 8% (n = 4) of patients, with a median of 21 days (IQR 20-30) from SXT initiation and led to discontinuation (n = 3). Conclusion SXT appears to be effective for treatment of BJIs as a salvage therapy, even in GNB or polymicrobial infection, including ODI. Further data are needed to confirm SXT efficacy as an alternative oral regimen in BJIs.

Perforations of surgical gloves are common and increase with the duration of glove wear. Skin flora, re-grown after pre-operative disinfection of the hands, may contaminate a surgical site. An antimicrobial surgical glove with chlorhexidine on its inner surface has been developed. We hypothesized that by suppressing the re-growth of skin flora during the complete course of a surgical procedure, antimicrobial gloves may reduce the risk of surgical site contamination in the event of an intra-operative glove breach.We conducted a randomized, double-blind, single-center trial, to measure any differences in the bacterial skin populations of surgeons' hands during surgical procedures done with antimicrobial and non-antimicrobial surgical gloves [ISRCTN71391952]. In this study, 25 pairs of gloves were retrieved from 14 surgeons who donned them randomly on their dominant or non-dominant hand. The number of bacteria retrieved from glove fluid was measured and expressed as colony forming units (CFU)/mL.The median cfu/mL of antimicrobial gloves was 0.00 (LQ: 0.00 CFU/mL; UQ: 0.00 cfu/mL), with a mean log10 cfu/mL=0.02 (range: 0.00-0.30). The median CFU/mL of non-antimicrobial gloves was 54.00 (LQ: 3.00 cfu/mL; UQ: 100.00 cfu/mL) with a mean log10 CFU/mL=1.32 (range: 0.00-2.39). After a mean operating time of 112 min, the difference in the log10 CFU/mL was 1.30 (p0.001).A new antimicrobial surgical glove suppressed surgeons' hand flora during operative procedures. In the event of a glove breach, the use of such a glove may have the potential to prevent bacterial contamination of a sterile surgical site, thereby decreasing the risk of surgical site infection (SSI) and increasing patient safety. Further clinical studies are needed to confirm this concept.

Glycopeptide-resistant Staphylococcus epidermidis (GRSE) strains are of increasing concern in bone and joint infections (BJIs). Using multilocus sequence typing and multilocus variable-number tandem repeat analysis, we show that BJI-associated GRSE strains are genetically diverse but arise from related, multiresistant hospital sequence types (STs), mostly ST2, ST5, and ST23.

We report a microbiological process for the documentation of prosthetic joint infection (PJI). Intraoperative periprosthetic tissue samples from 92 consecutive patients undergoing revision surgery for PJI were submitted to mechanized beadmill processing: specimens were aseptically collected in polypropylene vials, filled with sterile water and glass beads and submitted to mechanized agitation with a beadmill. The documentation rate of PJI following culture on solid and liquid media was 83.7% and the contamination rate 8.7%. Final documentation was obtained after overnight culture for 51.9% of cases and with 7 days of broth culture for all documented cases.

Summary Changes in the cell envelope composition of mycobacteria cause major changes in cytokine profiles of infected antigen presenting cells. We describe here the modulation of inflammatory responses by Mycobacterium abscessus, an emerging pathogen in cystic fibrosis. M. abscessus is able to switch from a smooth (S) to a rough (R) morphotype by the loss of a surface glycopeptidolipid. R variants are associated with severe clinical forms and a ‘hyper-proinflammatory’ response in ex vivo and in vivo models. Using partitioning of cell surface components we found that a complex fraction, more abundant in R variants than in S variants, made a major contribution to the TLR-2-dependent hyper-proinflammatory response induced by R variants. Lipoproteins were the main TLR-2 agonists in this fraction, consistent with the larger amounts of 16 lipoproteins in cell surface extracts from R variants; 15 out of 16 being more strongly induced in R variant than in S variant. Genetic interruption of glycopeptidolipid pathway in wild-type S variant resulted in R phenotype with similar induction of lipoprotein genes. In conclusion, R morphotype in M. abscessus is associated with increased synthesis/exposure at the cell surface of lipoproteins, these changes profoundly modifying the innate immune response through TLR-2-dependent mechanisms.

Introduction: Cefoxitin has a good in vitro activity and stability in resistance to hydrolysis by extended-spectrum beta-lactamases and is a good candidate for the treatment of urinary tract infection. However, data are scarce regarding its use in clinical practice. Methods: We conducted a retrospective study from September 2014 to November 2017, in a tertiary care hospital in Garches (France). We gathered all prescriptions of cefoxitin for urinary tract infection due to extended-spectrum beta-lactamase isolates. We compared the clinical outcomes between Escherichia coli and Klebsiella pneumoniae extended-spectrum-beta-lactamase-producing isolates after a 90-day follow-up. When available, we assessed whether cefoxitin-based regimen was associated with an emergence of resistance. Results: The treatment of 31 patients with a mean age of 60 ± 18 years was analyzed. We observed a clinical cure of 96.7% ( n = 30/31) at day 30 and of 81.2% ( n = 13/16) and 85.7% (12/14) at day 90 for extended-spectrum beta-lactamase Escherichia coli and Klebsiella pneumoniae isolates, respectively ( p = 0.72). No adverse events were reported. One patient who relapsed carried a Klebsiella pneumoniae isolate that became intermediate to cefoxitin in the follow-up. Conclusion: In a period of major threat with a continuous increase of extended-spectrum beta-lactamase obliging to a policy of carbapenem-sparing regimens, it seems detrimental to deprive physicians of using cefoxitin for extended-spectrum beta-lactamase Enterobacteriaceae for the treatment of urinary tract infection while our data show its efficacy.

A study was performed to compare matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS), linked to a recently engineered microbial identification database, and two rapid identification (ID) automated systems, BD Phoenix (Becton Dickinson Diagnostic Systems, France) and VITEK-2 (bioMérieux, Marcy L'Etoile, France), for the ID of coagulase-negative staphylococci (CoNS). Two hundred and thirty-four clinical isolates of CoNS representing 20 species were analyzed. All CoNS isolates were characterized by sodA gene sequencing, allowing interpretation of the ID results obtained using the respective database of each apparatus. Overall correct ID results were obtained in 93.2%, 75.6% and 75.2% of the cases with the MALDI-TOF-MS, Phoenix and VITEK-2 systems, respectively. Mis-ID and absence of results occurred in 1.7% and 5.1% of the cases with MALDI-TOF-MS, in 23.1% and 1.3% with the Phoenix, and in 13.7% and 0.9% with the VITEK-2 systems, respectively. In addition, with the latter automate, 10.3% of the IDs were proposed with remote possibility. When excluding the CoNS species not included in the databases of at least one of the three systems, the final percentage of correct results, Mis-ID and absence of ID were 97.4%, 1.3% and 1.3% with MALDI-TOF-MS, 79%, 21% and 0% with the Phoenix, and 78.6%, 10.3% and 0.9% with the VITEK-2 system, respectively. The present study demonstrates the robustness and high sensitivity of our microbial identification database used with MALDI-TOF-MS technology. This approach represents a powerful tool for the fast ID of clinical CoNS isolates.

Introduction La cefoxitine a une bonne activite in vitro et une stabilite face a l’hydrolyse des BLSE, lui permettant d’etre un bon candidat au traitement des infections urinaires (IU). Cependant, les donnees dans la litterature sont rares concernant son utilisation en pratique clinique. Materiels et methodes Etude retrospective de septembre 2014 a novembre 2017, dans un hopital universitaire (France). Nous avons rassemble toutes les prescriptions de cefoxitine pour des IU a BLSE, afin de comparer l’evolution clinique entre les souches de E. coli et de K. pneumonae a j90. Lorsque c’etait possible, nous avons egalement evalue si l’utilisation de la cefoxitine etait associee a une emergence de mutants resistants. Resultats Ont ete traites 31 patients avec un âge moyen de 60 ± 18 ans. Parmi eux, on note 22 pyelonephrites, 5 prostatites, 3 orchites et une cystite de l’homme. Nous avons observe une guerison clinique a j90 chez 81,2 % (n = 13/16) des cas pour les patients infectes a E. coli BLSE et de 85,7 % (12/14) pour les K. pneumoniae BLSE (p = 0,72). Soit une efficacite tout microorganisme confondu a 83,3 % (n = 25/30). La dose mediane de cefoxitine etait de 4 grammes par jour (de 2 a 8 grammes). Un seul patient infecte par un E. coli BLSE a ete relaye PO par levofloxacine. Aucun effet indesirable notable n’a ete rapporte dans le suivi. A noter qu’un patient a recidive avec un nouvel episode d’IU a K. pneumoniae devenu intermediaire a la cefoxitine apres avoir recu 6 grammes/j pour une prostatite et a justifie d’un nouveau traitement. Conclusion Ces donnees confirment l’efficacite de la cefoxitine dans le traitement des IU a BLSE. A ce jour, il s’agit de la cohorte la plus importante decrite comparativement a Kerneis et al. (n = 23) et Demonchy et al. (n = 23) demontrant 83 % de succes ainsi que Mambie et al. (n = 15) avec 9/10 analyses. Dans une periode d’augmentation croissante et certaine des BLSE, nous obligeant a appliquer une politique d’epargne des carbapenemes, il semble prejudiciable de se priver de l’utilisation de la cefoxitine en traitement des IU a BLSE, y compris pour des souches de K. pneumoniae, alors que nos donnees demontrent son efficacite.

Summary M. abscessus is the most common rapidly growing mycobacterium (RGM) pathogenic for man. It is by far the leading RGM causing lung infections in patients with an underlying disease (cystic fibrosis, COPD, bronchial dilatation), but also in healthy subjects free of lung disease. M. abscessus is also the most common RGM found in bronchial secretions in patients with cystic fibrosis. Besides their frequency, M. abscessus lung infections are generally true lung infections using the criteria established by the American Thoracic Society (ATS). The genome of the typical strain of M. abscessus has been sequenced and now provides means for molecular studies of the species with construction of deletion mutants. In addition, M. abscessus has a characteristic genotypic and phenotypic heterogeneity. M. abscessus, which was long considered to be a sub-species of M. chelonae, is now considered to be a complex including M. abscessus species stricto senso, M. bolletii and M. massiliense. Molecular identification being the only reliable identification method for M. abscessus, the study of different identification targets evidenced the mosaic makeup of some isolates, raising questions about the speciation process within the complex. Regarding the phenotype of M. abscessus sensu lato, two morphotypes can be described on solid media: the smooth (S) and the rough (R) morphotype. This heterogeneity is correlated with variable virulence, some isolates only being associated with true mycobacterial disease, these strains commonly displaying an R morphotype. The biochemical basis underlying the S and R phenotypes remain obscure, but recent studies shows that the R morphotype is associated with a small or highly diminished capacity for these bacteria to produce glycopeptidolipid (GPL) surface molecules. These recent data show that the R variants induce hyperproduction of TNF by murine macrophages in culture but also that the activation pathway leading to this process involves TLR2 receptors. The host response has been studied in mice. In this model, mycobacteria are typically controlled by the T cell compartment. An original humoral immune response has been demonstrated in the control of the emergence of rough strains. The study of the interaction between M. abscessus and other cell or animal models presenting a CFTR−/− or ΔF508 phenotype will be useful in understanding the specific sensitivity of children with cystic fibrosis to this mycobacterium.

Mycobacterium abscessusis an emerging rapidly growing mycobacterium that causes tuberculous-like lesions in humans. We studied the immune control of this organism in C57BL/6 mice challenged intravenously with 107CFU. Bacteria were eliminated from both the spleen and the liver within 90 days, and liver histology showed organized granulomatous lesions. A T- and B-cell requirement was investigated by challengingRag2−/−,Cd3ε−/−, and μMT−/−mice.Rag2−/−andCd3ε−/−mice were significantly impaired in the ability to clearM. abscessusfrom the liver and spleen, and μMT−/−mice were significantly impaired in the ability to clearM. abscessusfrom the liver, suggesting that infection control was primarily T cell dependent in the spleen and both T and B cell dependent in the liver. The liver granulomatous response was similar to that of wild-type controls in μMT−/−mice but completely absent inCd3ε−/−andRag2−/−mice. We studied the involvement of gamma interferon (IFN-γ) and tumor necrosis factor (TNF) by challenging C57BL/6 mice deficient in the IFN-γ receptor (Ifngr1−/−) and in TNF (Tnf−/−).Ifngr1−/−mice were significantly impaired inM. abscessuscontrol both in the spleen and in the liver, and granulomas were profoundly altered. The effect was even more substantial inTnf−/−mice; they failed to controlM. abscessusinfection in the liver and died within 20 to 25 days after infection with many hepatic inflammatory foci and major lesions of ischemic necrosis in the liver and kidney. These features were not observed with the closely related speciesM. chelonae. T-cell immunity, IFN-γ, and TNF are central factors for the control ofM. abscessusin C57BL/6 mice, as they are for the control of pathogenic slowly growing mycobacteria.

This article describes an outbreak of ACC-1-producing Klebsiella pneumoniae involving 40 patients. These were mainly men under 40 years old with a spinal cord injury, in a physical medicine and rehabilitation unit. The main risk factors were prolonged hospital stay, multiple-bed rooms, tracheostomy care and assisted defaecation. The outbreak was only controlled after the introduction of rigorous patient placement (i.e. single rooms or cohorting in the same room), while allowing the patients to have free access to the various technical services (e.g. physiotherapy and occupational therapy) and living spaces necessary for re-education.

The diagnosis and prognosis of lung infections due to the emerging nontuberculous mycobacterium (NTM) Mycobacterium abscessus are difficult to establish in children with cystic fibrosis.We evaluated the usefulness of an enzyme-linked immunosorbent assay for detecting serum IgG antibodies against the ubiquitous mycobacterial antigen A60.A total of 186 patients with cystic fibrosis (mean age+/-SD, 12.0+/-5.0 years) were studied, including 15 M. abscessus-positive patients who fulfilled American Thoracic Society (ATS) criteria for NTM infection (M. abscessus-infected patients), 7 M. abscessus-positive patients who did not fulfill ATS criteria, 20 patients positive for various other NTM who did not fulfill ATS criteria, and 144 NTM-negative control patients; mean IgG titers (+/-SD) for these 4 groups were 718+/-342 U, 193+/-113 U, 129+/-49 U, and 121+/-53 U, respectively (M. abscessus-infected patients vs. each of the other groups, P.005). The A60 IgG test was both sensitive (approximately 87%) and specific (approximately 95%) if adapted cutoff values were used (150 U and 250 U for patients agedor=10 years and patients aged10 years, respectively) and correlated well with results of acid-fast bacilli smears.Measurement of anti-A60 IgG may be useful for both the diagnosis and assessment of activity of M. abscessus lung infection in persons with cystic fibrosis.

Here we describe development of an extracorporeal hemoadsorption device for sepsis therapy that employs commercially available polysulfone or polyethersulfone hollow fiber filters similar to those used clinically for hemodialysis, covalently coated with a genetically engineered form of the human opsonin Mannose Binding Lectin linked to an Fc domain (FcMBL) that can cleanse a broad range of pathogens and endotoxin from flowing blood without having to first determine their identity. When tested with human whole blood in vitro, the FcMBL hemoadsorption filter (FcMBL-HF) produced efficient (90-99%) removal of Gram negative (Escherichia coli) and positive (Staphylococcus aureus) bacteria, fungi (Candida albicans) and lipopolysaccharide (LPS)-endotoxin. When tested in rats, extracorporeal therapy with the FcMBL-HF device reduced circulating pathogen and endotoxin levels by more than 99%, and prevented pathogen engraftment and inflammatory cell recruitment in the spleen, lung, liver and kidney when compared to controls. Studies in rats revealed that treatment with bacteriocidal antibiotics resulted in a major increase in the release of microbial fragments or 'pathogen-associated molecular patterns' (PAMPs) in vivo, and that these PAMPs were efficiently removed from blood within 2 h using the FcMBL-HF; in contrast, they remained at high levels in animals treated with antibiotics alone. Importantly, cleansing of PAMPs from the blood of antibiotic-treated animals with the FcMBL-hemoadsorbent device resulted in reduced organ pathogen and endotoxin loads, suppressed inflammatory responses, and resulted in more stable vital signs compared to treatment with antibiotics alone. As PAMPs trigger the cytokine cascades that lead to development of systemic inflammatory response syndrome and contribute to septic shock and death, co-administration of FcMBL-hemoadsorption with antibiotics could offer a more effective approach to sepsis therapy.

We report the case of a total hip arthroplasty infection caused by Ruminococcus gnavus in a 62-year-old man with ulcerative colitis. The bacterium was perfectly identified by matrix-assisted laser desorption ionization–time of flight mass spectrometry.

An atypical Enterobacteriaceae strain with a β-lactam susceptibility pattern of inducible cephalosporinase was isolated in Tenon Hospital (Paris, France) from a patient’s skull wound infection. Identifications by the API-50CHE biochemical system and 16S rRNA gene sequencing concluded that it was a member of the Buttiauxella genus. The bla gene was cloned and sequenced. The deduced translated product was a 383-amino acid protein (BUT-1) with 75–78% identity with the chromosomal AmpC β-lactamases of Citrobacter freundii , Enterobacter aerogenes , Enterobacter cloacae and Escherichia coli . The isoelectric point of 9.0 and the kinetic constants of BUT-1 were comparable with results described for other Ambler class C enzymes. bla BUT-1 and the associated ampR transcriptional regulator gene were divergently transcribed from a common intercistronic region, a genetic organization already described for other inducible class C β-lactamases. The deduced amino acid sequence of AmpR shared 85% and 81% identity with AmpR from E. cloacae and C. freundii respectively.

Introduction Le diagnostic preoperatoire d’infection de prothese osteoarticulaire (IPOA) est complexe, surtout dans les situations chroniques. De nombreux algorithmes sont utilises. Nous avons evalue la place de la serologie. Materiels et methodes Pendant une periode de 6 mois, nous avons inclus les patients prevus pour une revision de prothese de hanche ou de genou, en raison d’une suspicion d’IPOA chronique. La serologie etait prelevee en meme temps que les autres tests preoperatoires. Les resultats de la serologie etaient compares aux resultats histologiques et bacteriologiques peroperatoires. Les serologies etaient realisees avec le kit multiplex BJI Inoplex ® , qui utilise 16 antigenes recombinants diriges contre Staphylococcus species, Propionibacterium acnes et Streptococcus agalactiae . Resultats Quarante-cinq patients etaient inclus. La chirurgie precedente remontait a une mediane de 30 mois. La mediane de la CRP etait de 7 mg/L (extremes : 2 a 47) et de la vitesse de sedimentation de 12 mm (extremes : 2 a 50). Au final, le diagnostic d’IPOA etait retenu pour 12 patients et ecarte pour 33. Il s’agissait de staphylocoque coagulase negative (SCN) 5 fois, de P . acnes 4 fois, de candida sp. 2 fois et non defini 1 fois. La serologie etait concordante avec le diagnostic final pour 32 patients (25 steriles et 7 infectes). Pour 5 d’entre eux, la serologie fut le parametre essentiel du diagnostic preoperatoire. Dans ces cas, un SCN etait isole dans la ponction preoperatoire ou dans un seul des 5 prelevements peroperatoires ; la serologie negative 4 fois permettait de conclure a une contamination ; positive une fois, elle a fait poser le diagnostic d’IPOA. Dans cette etude, la serologie avait une sensibilite de 64 %, une specificite de 74 %, une valeur predictive positive de 44 % et une valeur predictive negative de 86 %. Dans le sous-groupe de 9 patients ayant une CRP > 10 mg/L, la serologie atteignait une sensibilite de 100 % avec la meme specificite. Conclusion Nous avons evalue la place de la serologie dans les situations les plus complexes de suspicion d’IPOA. Les resultats modestes peuvent s’expliquer par la nature des antigenes du test qui ne sont pas toujours ceux exprimes dans les bacteries du biofilm. Neanmoins ce test simple, combine aux autres parametres preoperatoires, peut apporter une aide utile au diagnostic.

Introduction La prise en charge des patients dans les Centres de reference des infections osteoarticulaires (CRIOA) permet de decider des traitements anti-infectieux lors des reunions de concertation pluridisciplinaire (RCP). Cependant ces strategies sont souvent complexes et potentiellement source d’erreurs lors de leur prescription. Nous nous sommes interesses a la frequence et aux types d’erreurs survenant lors de la prescription des anti-infectieux. Materiels et methodes Etude monocentrique prospective de novembre 2016 a janvier 2017 des erreurs lors de la saisie informatique des prescriptions d’anti-infectieux decidees en RCP, au sein de l’unite de chirurgie orthopedique septique (CRIOA). En effet, le pharmacien effectue le jour meme une verification de l’adequation des prescriptions realisees avec les decisions de la RCP. Sont verifiees les conformites de l’indication a l’antibiotherapie, des molecules prescrites, de leurs posologies, de leurs durees et de leurs voies d’administration. En cas de non-conformite avec la RCP, un message electronique est envoye au prescripteur pour correction. Nous avons releve les non-conformites aux decisions prises, les types d’erreurs, les signalements du pharmacien aux prescripteurs et leur impact. Resultats Sur 9 RCP, 61 patients ont beneficie d’une decision therapeutique concernant leur antibiotherapie. Au total, 16 (26 %) prescriptions comportaient au moins une non-adequation avec la decision prise. Pour 2 d’entre elles, cette non-adequation etait justifiee par un nouveau resultat microbiologique. Pour les 14 autres (23 %), 20 types d’erreurs ont ete releves : 6 poursuites d’antibiotherapies non justifiees, 4 schemas therapeutiques non respectes (dont 3 relais per os trop precoces), 4 antibiotherapies non arretees, 2 erreurs de posologie, 2 oublis de prescription d’un antibiotique, 1 prescription injustifiee d’un nouvel antibiotique et 1 erreur de voie d’administration. Les molecules les plus souvent concernees par ces erreurs etaient la rifampicine ( n = 7), la levofloxacine ( n = 4), l’amoxicilline/clavulanate ( n = 3) et la cefazoline ( n = 2). La lecture du message a permis la correction rapide des erreurs dans 8 cas (57 %). Pour les 6 autres prescriptions, l’intervention de l’infectiologue a ete necessaire pour permettre leur correction. Conclusion La participation du pharmacien clinicien aux RCP permet d’assurer le respect des decisions ainsi que de controler la prise en charge medicamenteuse des patients. Il constitue ainsi un element supplementaire de la securisation des prescriptions d’anti-infectieux.

A total of 212 coagulase-negative Staphylococcus strains recovered prospectively during 119 surgeries for proven or suspected bone and joint infection (BJI) were identified by sodA sequencing. These strains were identified as 151 Staphylococcus epidermidis isolates, 15 S. warneri isolates, 14 S. capitis isolates, 9 S. hominis isolates, 6 S. lugdunensis isolates, 5 S. haemolyticus isolates, 4 S. caprae isolates, 4 S. pasteuri isolates, 3 S. simulans isolates, and 1 S. cohnii isolate. Only S. epidermidis , S. lugdunensis , S. capitis , and S. caprae were found to be infecting organisms and were involved, respectively, in 35 (81.4%), 3 (7.0%), 3 (7.0%), and 2 (4.6%) cases of BJI.

Here we describe a blood-cleansing device for sepsis therapy inspired by the spleen, which can continuously remove pathogens and toxins from blood without first identifying the infectious agent. Blood flowing from an infected individual is mixed with magnetic nanobeads coated with an engineered human opsonin--mannose-binding lectin (MBL)--that captures a broad range of pathogens and toxins without activating complement factors or coagulation. Magnets pull the opsonin-bound pathogens and toxins from the blood; the cleansed blood is then returned back to the individual. The biospleen efficiently removes multiple Gram-negative and Gram-positive bacteria, fungi and endotoxins from whole human blood flowing through a single biospleen unit at up to 1.25 liters per h in vitro. In rats infected with Staphylococcus aureus or Escherichia coli, the biospleen cleared90% of bacteria from blood, reduced pathogen and immune cell infiltration in multiple organs and decreased inflammatory cytokine levels. In a model of endotoxemic shock, the biospleen increased survival rates after a 5-h treatment.

Here we describe how Staphylococcus aureus bacteria can be rapidly isolated from clinical samples of articular fluid and synovial tissue using magnetic beads coated with the engineered chimeric human opsonin protein, Fc-mannose-binding lectin (FcMBL). The FcMBL-beads were used to capture and magnetically remove bacteria from purified cultures of 12 S. aureus strains, and from 8 articular fluid samples and 4 synovial tissue samples collected from patients with osteoarthritis or periprosthetic infections previously documented by positive S. aureus cultures. While the capture efficiency was high (85%) with purified S. aureus strains grown in vitro, direct FcMBL-bead capture from the clinical samples was initially disappointing (< 5% efficiency). Further analysis revealed that inhibition of FcMBL binding was due to coating of the bacteria by immunoglobulins and immune cells that masked FcMBL binding sites, and to the high viscosity of these complex biological samples. Importantly, capture of pathogens using the FcMBL-beads was increased to 76% efficiency by pretreating clinical specimens with hypotonic washes, hyaluronidase and a protease cocktail. Using this approach, S. aureus bacteria could be isolated from infected osteoarthritic tissues within 2 hours after sample collection. This FcMBL-enabled magnetic method for rapid capture and concentration of pathogens from clinical samples could be integrated upstream of current processes used in clinical microbiology laboratories to identify pathogens and perform antibiotic sensitivity testing when bacterial culture is not possible or before colonies can be detected.

L’infection osseuse est une affection grave aux etiologies et mecanismes physiopathologiques multiples. Sous le terme largement employe d’osteite ou d’osteomyelite (les Anglo-Saxons utilisant quasiment exclusivement le terme imprecis de osteomyelitis), les series de la litterature rassemblent plusieurs entites pathologiques assez distinctes, allant de l’infection osseuse sur os solide (ou continu) a l’infection sur prothese articulaire, en passant par l’osteomyelite hematogene et la pseudarthrose suppuree, et il n’y a pas a l’heure actuelle de definition claire de l’osteite. Nous preferons parler d’infection osseuse sur os continu. Ainsi, dans cette situation, le probleme n’est pas d’obtenir la consolidation osseuse ou de savoir quand pouvoir reposer une prothese articulaire. Lors d’une infection sur os solide, les difficultes sont de pouvoir combler une cavite osseuse, d’avoir une antibiotherapie efficace au niveau osseux, et d’obtenir une bonne couverture cutanee. Ainsi, dans notre pratique, le schema therapeutique de toute infection sur os continu est stereotype en trois temps: excision de tous les tissus infectes (parties molles et os) ou a vitalite douteuse; comblement de la cavite osseuse residuelle; couverture cutanee par des parties molles bien vascularisees.

We studied 315 coagulase-negative Staphylococcus strains recovered prospectively during 240 surgical procedures (206 subjects) from proven or suspected device-associated bone and joint infections. Sixteen strains (5.1%) had decreased susceptibility to glycopeptides: 15 (12 S. epidermidis strains, 2 S. capitis strains, and 1 S. haemolyticus strain) to teicoplanin alone (MIC of 16 mg/liter, n = 9; MIC of 32 mg/liter, n = 6) and one ( S. epidermidis ) to both teicoplanin and vancomycin (MIC, 16 and 8 mg/liter, respectively). Decreased susceptibility to teicoplanin was more prevalent in “infecting” strains (i.e., strains recovered from ≥2 distinct intraoperative samples) than in “contaminants” (i.e., strains not fulfilling this criterion) (8.1% [12/149] versus 2.4% [4/166], respectively [ P = 0.022]). One hundred percent (13/13) of S. epidermidis strains with decreased susceptibility to teicoplanin were resistant to methicillin (versus 112/173 [64.7%] for S. epidermidis strains susceptible to teicoplanin; P = 0.021).

We performed a multicenter prevalence study of nontuberculous mycobacteria (NTM) involving 1,582 patients (mean age, 18.9 years; male/female ratio, 1.06) with cystic fibrosis in France. The overall NTM prevalence (percentage of patients with at least one positive culture) was 6.6% (104/1,582 patients), with prevalences ranging from 3.7% (in the east of France) to 9.6% (in the greater Paris area). Mycobacterium abscessus complex (MABSC; 50 patients) and Mycobacterium avium complex (MAC; 23 patients) species were the most common NTM, and the only ones associated with fulfillment of the American Thoracic Society bacteriological criteria for NTM lung disease. The "new" species, Mycobacterium bolletii and Mycobacterium massiliense, accounted for 40% of MABSC isolates. MABSC species were isolated at all ages, with a prevalence peak between 11 and 15 years of age (5.8%), while MAC species reached their highest prevalence value among patients over 25 years of age (2.2%). Nontuberculous mycobacteria (NTM) have emerged as "new" pathogens in cystic fibrosis (CF) patients over the last 2 decades (10). CF centers worldwide have reported isolation of NTM from the respiratory tracts of CF patients, with prevalence values ranging from 5% to 20% (5, 6, 8, 9, 13, 14, 16, 19, 22, 25). Mycobacterium avium complex (MAC) and Mycobac-terium abscessus complex (MABSC) species are the most frequently isolated NTM and together account for 95% of NTM lung diseases affecting CF patients. The MAC, a member of the subgroup comprising slowly growing mycobacteria, ranks first in North America (22), whereas the MABSC, a member of the subgroup comprising rapidly growing mycobacteria, seems to predominate in Western Europe (15, 23, 25) and is also more prevalent than the MAC in Israel (19). Previous studies have reported isolation of NTM from 6.6 to 9.8% of French CF cohorts (9, 23, 25). These studies also reported a much higher isolation rate for MABSC than for MAC or other NTM species (23, 25). However, these studies were done exclusively in pediatric CF centers in Paris. This may have distorted the results since MABSC species are more prevalent than MAC species in children (23). Moreover, the epidemiology of NTM in Paris does not necessarily reflect the situation in other regions of France. For example, studies involving non-CF patients have reported higher rates of NTM disease in urban areas (20). Moreover, previous French studies were performed before M. abscessus (now M. abscessus sensu lato, or the MABSC) was shown to include at least three distinct species, M. abscessus (sensu stricto) (hereafter referred to as M. abscessus), Mycobacterium massiliense, and Mycobac-terium bolletii (1, 3). The prevalences of these three species in CF patients in France were therefore unknown. We thus conducted a large, prospective, nationwide study addressing NTM prevalence in CF patients in France. This study shows relatively low prevalence figures for French CF centers. It also provides evidence that MABSC species are currently the most prevalent NTM in the French CF population , far more frequent than MAC species, and that MABSC species mainly infect children and young adults. The 49 French CF centers were asked to participate in the study. Each patient included was asked to provide at least three sputum samples (or other respiratory specimens) for NTM analysis between 1 January 2004 and 31 December 2004. Prior NTM isolation did not exclude subjects. Patients with a positive NTM sample had to submit three more sputum samples at monthly intervals and then every 3 months for a total of 18 months. All the patients, or their parents if they were chil-* Corresponding author. Mailing address: Laboratoire de Microbi-ologie, Hôpital Ambroise Paré

International audience; Propionibacterium acnes is the most frequent anaerobic pathogen found in spondylodiscitis. A documented case required microbiological proof of P. acnes with clinical and radiological confirmation of inflammation in a localized region of the spine. Microbiological samplings were obtained by surgery or aspiration under radiological control. Twelve males and 17 females (median age, 42 years) with spondylodiscitis due to P. acnes were diagnosed within the last 15 years. Three patients were immunosuppressed. All patients reported back pain as the main symptom, and most were afebrile. Three patients had a peripheral neurological deficit, one a motor deficit, and two a sensory deficit attributable to the infection; and six patients had an epidural abscess. The most frequent risk factor was surgery, which was present in the history 28 of 29 (97%) patients. The mean delay between spinal surgery and onset of disease was 34 months, with a wide range of 0-156 months. Osteosynthesis material was present in twenty-two cases (76%). In 24 (83%) patients, additional surgery, such as débridement or spondylodesis, was performed. Previous osteosynthesis material was removed in 17 of the 22 (77%) patients where it was present. Total cure was reported in all patients, except one, after a mean duration of antibiotic therapy of 10.5 weeks (range, 2-28 weeks). In conclusion, spondylodiscitis due to P. acnes is an acute infection closely related to previous surgery. The most prominent clinical feature is pain, whereas fever is rare, and the prognosis is very good.