Evaluation of the MRSA/SA ELITe MGB Assay for the Detection of Staphylococcus aureus in Bone and Joint Infections.

Bone and joint infections represent a potentially devastating complication of prosthetic orthopedic joint replacement, thus requiring both rapid and appropriate antibiotic treatment. Staphylococcus aureus is one of the most common pathogens involved in this pathology. Being able to assert its presence is the first step of efficient patient management. This monocenter study evaluated the MRSA/SA ELITe MGB assay for the molecular detection of S. aureus and methicillin-resistant S. aureus (MRSA) in bone and joint biopsy specimens and synovial fluids. This test, together with conventional techniques, including standard cultures and the 16S rRNA amplification assay, was performed on 208 successive perioperative samples collected prospectively for 1 year obtained from 129 patients. Using conventional techniques, we detected a microbial pathogen in 76 samples from 58 patients, 40 of which were identified as S. aureus. The limit of detection (LOD) of the MRSA/SA ELITe MGB assay was experimentally determined for bone and joint biopsy specimens and synovial fluids using negative samples spiked with S. aureus ATCC 43300. The sensitivities of S. aureus detection with the MRSA/SA ELITe MGB assay were 82.5% (33/40 samples) and 97.5% (39/40 samples) using the manufacturer's LOD and an experimentally determined LOD, respectively. Interestingly, using the osteoarticular specific LOD, 15 additional samples were determined to be positive for S. aureus DNA with the MRSA/SA ELITe MGB assay; in all cases, these samples were obtained from patients considered to be infected with S. aureus according to their clinical and microbiological records. The results were available within 24 h, which could help to expedite therapeutic decisions.

Intracellular activity of antimicrobial compounds used for Staphylococcus aureus nasal decolonization.

Background: Staphylococcus aureus is able to invade mammalian cells during infection and was recently observed inside nasal mucosa of healthy carriers. Objectives: To determine the intracellular activity of antimicrobial compounds used for decolonization procedures using a cell model mimicking S. aureus nasal epithelium invasion. Patients and methods: HaCaT cells and human nasal epithelial cells (HNECs) recovered from nasal swabs of S. aureus carriers were visualized by confocal laser scanning microscopy to detect intracellular S. aureus cells. An HaCaT cell model, mimicking S. aureus internalization observed ex vivo in HNECs, was used to assess the intracellular activity against S. aureus of 21 antimicrobial compounds used for nasal decolonization, including mupirocin and chlorhexidine. Results: HaCaT cells and HNECs were found to internalize S. aureus with the same focal pattern. Most antimicrobial compounds tested on HaCaT cells were shown to have weak activity against intracellular S. aureus. Some systemic antimicrobials, including fusidic acid, clindamycin, linezolid, minocycline, ciprofloxacin, moxifloxacin, rifampicin and levofloxacin, reduced S. aureus intracellular loads by 0.43-1.66 log cfu/106 cells compared with the control (P < 0.001). By contrast, mupirocin and chlorhexidine reduced the S. aureus intracellular load by 0.19 and 0.23 log cfu/106 cells, respectively. Conclusions: These data indicate that most of the antimicrobial compounds used for nasal decolonization, including mupirocin and chlorhexidine, exhibit weak activity against intracellular S. aureus using the HaCaT cell model. This work emphasizes the need to better understand the role of the S. aureus intracellular reservoir during nasal colonization in order to improve decolonization procedures.

Carbapenemase-producing Acinetobacter baumannii: An outbreak report with special highlights on economic burden.

OBJECTIVE: We aimed to describe the management of a carbapenemase-producing Acinetobacter baumannii (CP-AB) outbreak using the Outbreak Reports and Intervention Studies of Nosocomial Infection (ORION) statement. We also aimed to evaluate the cost of the outbreak and simulate costs if a dedicated unit to manage such outbreak had been set-up. METHODS: We performed a prospective epidemiological study. Multiple interventions were implemented including cohorting measures and limitation of admissions. Cost estimation was performed using administrative local data. RESULTS: Five patients were colonized with CP-AB and hospitalized in the neurosurgery ward. The index case was a patient who had been previously hospitalized in Portugal. Four secondary colonized patients were further observed within the unit. The strains of A. baumannii were shown to belong to the same clone and all of them produced an OXA-23 carbapenemase. The closure of the ward associated with the discharge of the five patients in a cohorting area of the Infectious Diseases Unit with dedicated staff put a stop to the outbreak. The estimated cost of this 17-week outbreak was $474,474. If patients had been managed in a dedicated unit - including specific area for cohorting of patients and dedicated staff - at the beginning of the outbreak, the estimated cost would have been $189,046. CONCLUSION: Controlling hospital outbreaks involving multidrug-resistant bacteria requires a rapid cohorting of patients. Using simulation, we highlighted cost gain when using a dedicated cohorting unit strategy for such an outbreak.

Correlation between clinical data and antibiotic resistance in coagulase-negative Staphylococcus species isolated from 68 patients with acute post-cataract endophthalmitis.

Coagulase-negative staphylococci (CNS) cause the majority of post-cataract endophthalmitis, which can lead to anatomical and/or functional loss of the eye. This study reports the antibiotic susceptibilities of CNS isolates associated with acute post-cataract endophthalmitis cases and correlates antibiotic resistance with severity and outcome of infection in these patients. Clinical data (initial ocular examination, final prognosis, antibiotic treatment) and the antibiotic susceptibilities of the isolated CNS strains were obtained from 68 patients with post-surgical endophthalmitis recruited during a 7-year period by the FRench Institutional ENDophthalmitis Study (FRIENDS) group. The CNS strains displayed 100% susceptibility to vancomycin, 70% to fluoroquinolones, 83% to fosfomycin, 46% to imipenem and 18% to piperacillin. The most effective antibiotic combinations were fosfomycin plus a fluoroquinolone and imipenem plus a fluoroquinolone, which were considered adequate in 80% and 58% of patients, respectively. Methicillin resistance was significantly associated with older age (p 0.001), diabetes mellitus (p 0.004), absence of fundus visibility (p 0.06), and poor visual prognosis (p 0.03). Resistance to fluoroquinolones was significantly associated with absence of fundus visibility (p 0.05) and diabetes mellitus (p 0.02). This large prospective study demonstrates that methicillin resistance and, to a lesser extent, fluoroquinolone resistance in CNS strains causing postoperative endophthalmitis are both prevalent in France and associated with a poorer visual prognosis. These results emphasize the need for an effective surveillance of this antibiotic resistance and the development of new diagnostic tools for rapid detection for early optimization of antibiotic therapy in endophthalmitis patients.

Risk factors for quinolone-resistance in women presenting with Escherichia coli acute pyelonephritis.

UNLABELLED: In France, according to the National Epidemiology Observatory of Bacterial Resistance to Antibiotics, 15.3% of outpatient urinary Escherichia coli isolates were fluoroquinolone-resistant in 2010. This puts to question the relevance of empirical fluoroquinolone therapy for community-acquired acute pyelonephritis (APN), potentially severe infections. OBJECTIVES: We had for aim to identify individual risk factors for quinolone-resistant E. coli in community-acquired APN. PATIENTS AND METHODS: A retrospective cohort study of 344 adult female patients presenting with E. coli APN was conducted at the Roanne and Saint-Etienne hospital emergency departments, from January 2011 to February 2012. We studied the demographic, administrative, and clinical factors. E. coli strains with intermediate susceptibility on the antibiogram were considered as resistant. RESULTS: There was 23% of isolates that were resistant to nalidixic acid and 17.4% to ofloxacin. Complicated APN was not a significant risk factor (univariate analysis). Three risk factors of resistance to nalidixic acid and ofloxacin were independent (multivariate analysis): fluoroquinolone use in the previous 3 months, hospitalization in the previous 6 months, and stay in a long-term care facility. The resistance to ofloxacin reached 30.6% if at least 1 of these risk factors was present; it was 9% when none of the factors were present. CONCLUSIONS: These results suggest that local recommendations for the empirical therapy of APN should be reviewed. The limitations of our study require backing up our results with prospective multicentric studies that could lead to drafting new national recommendations.

Nosocomial transmission of NDM-1-producing Escherichia coli within a non-endemic area in France.

Two patients with no travel history and sharing the same room were colonized by the same strain of New Delhi metallo-ß-lactamase 1 (NDM-1)-producing Escherichia coli within a geographical area not endemic for this highly multidrug-resistant bacterium. It was documented an absence of an epidemiological and bacteriological link with a third patient returning from India after surgery and found to be infected by an NDM-1-producing Citrobacter strain during the same period. Despite extensive investigation, the source of contamination of the two former patients was not elucidated. This case report illustrates the need of investigating rapidly the emergence of highly multidrug-resistant Enterobacteriaceae, to stop their dissemination in a nosocomial setting.

An algorithm based on one or two nasal samples is accurate to identify persistent nasal carriers of Staphylococcus aureus.

Persistent Staphylococcus aureus nasal carriers are at high risk of S. aureus infection. The present study delineates a simple strategy aimed at identifying rapidly and accurately this subset of subjects for clinical or epidemiological purposes. Ninety healthy volunteers were each identified as persistent, intermittent or non-nasal carriers of S. aureus by using seven specimens sampled over a 5-week period. By reference to this so-called reference standard, six other strategies aimed at simplifying and speeding the identification of persistent carriers and based on the qualitative or quantitative detection of S. aureus in one to three nasal samples were evaluated by the measure of the area under the curve of receiver operating characteristic diagrams. Among strategies using qualitative results, there was no statistical difference between protocols using seven and three samples. A threshold of 10(3) CFU of S. aureus per swab was found capable of defining persistent nasal carriage with a sensitivity of 83.1% and a specificity of 95.6%. These figures reached 95.5% and 94.9%, respectively, by using an algorithm including one or two nasal specimens according to the threshold of 10(3) CFU of S. aureus in the first swab. The latter two strategies were shown to be costly equivalents. The proposed algorithm-based strategy proved to be relevant to identify properly and consistently persistent nasal carriers of S. aureus. However, as it was built from data of healthy volunteers, it needs to be confirmed prospectively on patients potentially at risk for S. aureus infection.

[Dosage adjustment of vancomycin in continuous infusion in critically ill-patients]. / Ajustement de la posologie de la vancomycine administrée en perfusion continue chez des patients de réanimation.

INTRODUCTION: As the susceptibility of staphylococcal strains to glycopeptides rises, it is becoming necessary to increase vancomycin dosages. OBJECTIVE: To evaluate an administration protocol for vancomycin using continuous infusion with a loading dose of 30 mg/kg followed by 30 mg/kg per 24h in intensive care patients presenting creatinine clearance (CLc) greater than 50. RESULTS: A total of 22 patients were included in the study. Serum vancomycin concentrations after 24h (C24h) ranged from 25 to 30 mg/l in seven of 14 patients with CLc less than 120 ml/min (50 %), compared with three patients (21 %) with C24h greater than 35 mg/l and four patients (29 %) with C24 h less than 25 mg/l. However, C24h was less than 20mg/l for the eight patients with CLc greater or equal to 120 ml/min. Bacteriological data was available for eight of the 14 patients with CLc less than 120 ml/min, and in these eight patients, the C24h/MIC was greater or equal to 8; seven of these patients had an AUC/MIC greater or equal to 350. CONCLUSION: Assay of serum vancomycin concentrations after 24h of treatment is necessary to enable rapid adjustment of vancomycin concentration in order to improve therapeutic efficacy or avoid nephrotoxicity.

Optimization of the use of ciprofloxacin.

AIMS: To compare mutant prevention concentration (MPC) of ciprofloxacin and time-killing curve with regards to 11 genotyped Escherichia coli. METHOD: MICs were determined using the E-test method. Time-killing studies were performed in accordance with the NCCLS guidelines. The genes gyrA, gyrB, parC, parE and marR were amplified by PCR and sequenced. The MPC was defined as the lowest antibiotic concentration preventing the growth of resistant colonies when 10(10) CFU/mL were spread on a solid medium. RESULTS: Strains with no genes gyrA, gyrB, parC, parE and marR mutation presented MIC less or equal to 0.023 mg/L and MPC less or equal to 0.25 mg/L. Strains with two mutations (gyrA and parC) presented MIC equal to 1.5 mg/L and MPC equal to 4 mg/L. Strains with one mutation (gyrA) presented MIC less or equal to 0.75 mg/L, but MPC ranged from 0.5 to 6 mg/L depending of the MIC of ciprofloxacin. The time-killing curves for ciprofloxacin showed a bactericidal activity of 0.25 mg/L in 1h for strains without mutation, compared with a bactericidal activity of 2 and 4 mg/L in 4h for strains with one and two mutations, respectively. CONCLUSION: For strains of E. coli resistant to nalidixic acid, it was necessary to evaluate the MIC of ciprofloxacin in order to asses the optimal dosage of ciprofloxacin.

Comparison of bacteriostatic and bactericidal activity of 13 essential oils against strains with varying sensitivity to antibiotics.

AIMS: To compare the bacteriostatic and bactericidal activity of 13 chemotyped essential oils (EO) on 65 bacteria with varying sensitivity to antibiotics. METHODS AND RESULTS: Fifty-five bacterial strains were tested with two methods used for evaluation of antimicrobial activity (CLSI recommendations): the agar dilution method and the time-killing curve method. EO containing aldehydes (Cinnamomum verum bark and Cymbopogon citratus), phenols (Origanum compactum, Trachyspermum ammi, Thymus satureioides, Eugenia caryophyllus and Cinnamomum verum leaf) showed the highest antimicrobial activity with minimum inhibitory concentration (MIC) <2% (v/v) against all strains except Pseudomonas aeruginosa. Alcohol-based EO (Melaleuca alternifolia, Cymbopogon martinii and Lavandula angustifolia) exhibited varying degrees of activity depending on Gram status. EO containing 1.8-cineole and hydrocarbons (Eucalyptus globulus, Melaleuca cajeputii and Citrus sinensis) had MIC(90%) > or = 10% (v/v). Against P. aeruginosa, only C. verum bark and O. compactum presented MIC < or =2% (v/v). Cinnamomum verum bark, O. compactum, T. satureioides, C. verum leaf and M. alternifolia were bactericidal against Staphylococcus aureus and Escherichia coli at concentrations ranging from to 0.31% to 10% (v/v) after 1 h of contact. Cinnamomum verum bark and O. compactum were bactericidal against P. aeruginosa within 5 min at concentrations <2% (v/v). CONCLUSIONS: Cinnamomum verum bark had the highest antimicrobial activity, particularly against resistant strains. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacteriostatic and bactericidal activity of EO on nosocomial antibiotic-resistant strains.

Propionibacterium acnes contamination in lumbar disc surgery.

Previous studies have shown that Propionibacterium acnes may be responsible for low-grade infection of the intervertebral discs of patients with severe sciatica. The aim of this study was to prospectively investigate the presence of bacteria in disc fragment samples obtained during surgery for lumbar disc herniation. P. acnes was cultured from disc fragments in two (3.7%) of 54 patients studied. In addition, control cultures taken from ligamentum flavum and muscle from these two patients were also positive for P. acnes. Similar control cultures were positive for P. acnes from a further ten (18.5%) patients. Four air samples taken during surgery all contained P. acnes; the organism was also found from three of 54 laminar flow control cultures. Sample contamination appears the most likely cause for the presence of P. acnes in the lumbar disc fragment cultures.

Outbreak of postoperative shoulder arthritis due to Propionibacterium acnes infection in nondebilitated patients.

We investigated an outbreak of postoperative shoulder arthritis due to Propionibacterium acnes infection in nondebilitated patients. Risk factors were male sex, the order in which surgery was performed during the daily operating schedule, and increased duration of the surgical procedure. After modification of the ventilation system and implementation of improved cleaning methods in the operating theater, no new cases were recorded.

Arthroplastic and osteosynthetic infections due to Propionibacterium acnes: a retrospective study of 52 cases, 1995-2002.

The cases of 52 patients with Propionibacterium acnes infection of orthopaedic implants are summarized: 20 patients with definite infection (sepsis, with P. acnes recovered from multiple specimens per patient), 15 with probable infection (sepsis, with P. acnes recovered from one specimen), and 17 with possible infection (signs of prosthetic malfunction or pseudo-osteoarthritis, with P. acnes recovered from one specimen). The patient population consisted of 37 males and 15 females with a mean age of 51.8 years (range 17-88). Besides bone surgery, 21% of these patients had severe coexisting illness. The study population was very heterogeneous and clinical presentation very polymorphic; infections became clinically apparent through sepsis, prosthetic malfunction, or a delay in consolidation. The diagnosis was highly dependent on the quality of the samples taken and the methodology used by the microbiology laboratory to isolate this bacterium. Culture time was long, on average 11.4 days. Treatment involved a combination of antibiotic treatments (67% of cases) and ablation of the material (83% of cases). Although P. acnes is considered to be weakly pathogenic, this bacterium may be responsible for infections in patients with implanted orthopaedic material. Ablation of the arthroplastic or osteosynthetic material is necessary in the majority of cases.

Efficiency of blood culture bottles for the fungal sterility testing of corneal organ culture media.

BACKGROUND/AIM: The consequences of fungal contamination of an organ cultured cornea, though exceptional, are often disastrous for the recipient. Consequently, eye banks often quarantine corneas for 10 days or more before passing them for grafting. This period, though detrimental to the endothelial cell density of the delivered cornea, is necessary to detect contamination using conventional microbiological methods. The authors previously validated the use of a pair of aerobic and anaerobic blood bottles for sensitive and rapid detection of bacteria. To allow a short quarantine period, it remained only to optimise detection of fungi. The authors aimed to compare sensitivity and rapidity of fungal contamination detection by three methods: blood bottles, Sabouraud, and daily visual inspection of the organ culture medium. METHODS: Four inocula (10(6), 10(4), 10(2), 10 colony forming unit (CFU) per ml) of 11 fungi (Candida albicans, C tropicalis, C glabrata, Saccharomyces cerevisiae, Rhodotorula rubra, Cryptococcus neoformans, Fusarium oxysporum, Aspergillus niger, A fumigatus, A flavus, Acremonium falciforme) were inoculated in a commercial organ culture medium containing a coloured pH indicator (CorneaMax, Eurobio, Les Ulis, France). The real live fungal inoculum was verified immediately after inoculation. After 48 hours at 31 degrees C, samples of the contaminated media were inoculated in three blood bottles: Bactec Aerobic/F, Bactec Mycosis IC/F, and Bactec Myco/F Lytic (Becton Dickinson, Le Pont de Claix, France), then placed in a Bactec 9240 rocking automat, and in four Sabouraud media (solid and liquid, 28 degrees C and 37 degrees C) with daily observation. Contaminated organ culture media were also checked daily for any change in turbidity and/or colour. Experiments were performed in triplicate. RESULTS: Mycosis IC/F and Myco/F Lytic bottles were neither faster nor more sensitive than the aerobic bottle. The three methods were positive for all inocula, even the lowest (viable inoculum below 10 CFU/ml for each fungus). Contamination was detected within 24 hours by the aerobic bottles in 91% (40/44), by Sabouraud in 98% (43/44) (no significant difference) and by visual inspection in 66% of cases (29/44) (p<0.001 with the two others). Maximum times to detection were 46, 48 and 72 hours respectively. CONCLUSION: This study further counters the preconception that fungal contamination is hard to detect in corneal organ culture media. This study is the last step in validating the use of a pair of blood bottles for the sterility testing of organ culture media, this time for fungi. Their use should make it possible to shorten microbiological quarantine and thus deliver corneas with higher endothelial cell density, without increasing the risk of recipient contamination.

Impact of restricting fluoroquinolone prescription on bacterial resistance in an intensive care unit.

The purpose of this study was to assess the effect of reducing prescription of fluoroquinolones in an intensive care unit (ICU) upon bacterial resistance, particularly as regards Pseudomonas aeruginosa. For six months between January 2001 and June 2001, administration of fluoroquinolones was kept to a minimum. A bacteriological screening of patients was performed to assess the incidence of fluoroquinolone-resistant bacteria. There was a 75.8% restriction in prescriptions of fluoroquinolones. There was no significant change in bacterial ecology between the periods preceding (12 months) and following (12 months) restriction. There was a significant recovery of sensitivity of P. aeruginosa to ciprofloxacin (P

[Medical-surgical treatment of pulmonary infection with Mycobacterium malmoense]. / Infection pulmonaire à Mycobacterium malmoense, traitement médico-chirurgical.

CASE REPORT: We report the case of a patient with chronic obstructive pulmonary disease in whom pulmonary infection due to mycobacterium malmoense was discovered unexpectedly. A diagnostic and therapeutic surgical resection was performed. CONCLUSION: The non-tuberculous mycobacterium was identified by culture of the specimen. Surgery was followed by empirical antibiotic treatment with rifampicin and pyrazinamide for two and a half months. Isoniazid was withdrawn rapidly on account of hepatitis and the treatment was supplemented later with clarithromycin, leading to a total duration of treatment of seven and a half months. This case is unusual because of its medico-surgical management that led to assessment and appropriate treatment of this infection.

[Monitoring of antibiotic treatment of patient with a severe bacterial infection]. / Place du laboratoire dans le choix et le suivi pharmacodynamique de l'antibiothérapie des infections sévères.

OBJECTIVES: To provide a summary of useful up-to-date knowledge regarding experimental and clinical bacteriology, pharmacokinetics and pharmacodynamics in order to optimise efficacy of antibiotic treatment of hospital patients with serious bacterial infections. DATA SOURCES: Record of references from national and international journals in Medline. STUDY SELECTION: Extraction of the most relevant theoretical and practical data from studies published over the last 5 years. DATA SYNTHESIS: Changes in resistance to antibiotics, as well as the limited number of new antibacterial drugs available and the cost of therapeutic failure all militate in favour of a more elaborate approach to therapeutic strategies involving antibiotics, particularly regarding hospitalised patients. The efficacy of antibiotic therapy can be optimised through the utilization of bacteriological, pharmacokinetic and pharmacodynamic data, thereby increasing the likelihood of a successful outcome. While the antibiogram constitutes the fundamental analytical tool for evaluating the activity of antibiotics, the minimum inhibitory concentration (MIC) is of value in selecting appropriate drugs and dosages, particularly for bacterial strains having lower susceptibility. Screening for genes of resistance to antibiotics provides more accurate analysis of bacterial resistance. In recent years, the efficacy of antibiotics has been improved through the use of a number of pharmacodynamic parameters: inhibitory quotient (IQ), area under the serum concentration-time curve to MIC ratio (AUC/MIC) and the time the serum concentration is greater than the MIC (T > MIC). In standard practice, data readily available to the clinician comprise the MIC and serum antibiotic concentrations. There is some discussion concerning optimisation of antibiotic efficacy through the use of these parameters. CONCLUSION: Close collaboration between clinicians and microbiologists results in improved quality of antibiotic therapy and better management of antibiotics.

Detection of an outbreak of methicillin-resistant Staphylococcus aureus with reduced susceptibility to glycopeptides in a French hospital.

Staphylococcus aureus isolates were screened for reduced susceptibility to glycopeptides with an initial glycopeptide agar screening test, followed by confirmation of the strains thus identified by two Etest strip techniques and population analysis. This procedure detected 48 methicillin-resistant S. aureus (MRSA) isolates with reduced susceptibility to glycopeptides from 24 patients among 883 MRSA isolates tested. The dissemination of a single clone was confirmed by pulsed-field gel electrophoresis.