Stéphane Corvec

Genetics and Expression of the Carbapenem-Hydrolyzing Oxacillinase Gene blaOXA-23 in Acinetobacter baumannii

ABSTRACT The genetic structures surrounding the plasmid-carried blaOXA-23 oxacillinase gene, encoding resistance to carbapenems, were studied in Acinetobacter baumannii. ISAba1 and the novel element ISAba4 were detected upstream of the blaOXA-23 gene, providing promoter sequences for its expression. These insertion elements were likely involved in transposition processes at the origin of acquisition of this β-lactamase gene.

Propionibacterium acnes: An Underestimated Pathogen in Implant-Associated Infections

The role of Propionibacterium acnes in acne and in a wide range of inflammatory diseases is well established. However, P. acnes is also responsible for infections involving implants. Prolonged aerobic and anaerobic agar cultures for 14 days and broth cultures increase the detection rate. In this paper, we review the pathogenic role of P. acnes in implant-associated infections such as prosthetic joints, cardiac devices, breast implants, intraocular lenses, neurosurgical devices, and spine implants. The management of severe infections caused by P. acnes involves a combination of antimicrobial and surgical treatment (often removal of the device). Intravenous penicillin G and ceftriaxone are the first choice for serious infections, with vancomycin and daptomycin as alternatives, and amoxicillin, rifampicin, clindamycin, tetracycline, and levofloxacin for oral treatment. Sonication of explanted prosthetic material improves the diagnosis of implant-associated infections. Molecular methods may further increase the sensitivity of P. acnes detection. Coating of implants with antimicrobial substances could avoid or limit colonization of the surface and thereby reduce the risk of biofilm formation during severe infections. Our understanding of the role of P. acnes in human diseases will likely continue to increase as new associations and pathogenic mechanisms are discovered.

Epidemiology and new developments in the diagnosis of prosthetic joint infection

Although prosthetic joint infection (PJI) is a rare event after arthroplasty, it represents a significant complication that is associated with high morbidity, need for complex treatment, and substantial healthcare costs. An accurate and rapid diagnosis of PJI is crucial for treatment success. Current diagnostic methods in PJI are insufficient with 10–30% false-negative cultures. Consequently, there is a need for research and development into new methods aimed at improving diagnostic accuracy and speed of detection. In this article, we review available conventional diagnostic methods for the diagnosis of PJI (laboratory markers, histopathology, synovial fluid and periprosthetic tissue cultures), new diagnostic methods (sonication of implants, specific and multiplex PCR, mass spectrometry) and innovative techniques under development (new laboratory markers, microcalorimetry, electrical method, reverse transcription [RT]-PCR, fluorescence in situ hybridization [FISH], biofilm microscopy, microarray identification, and serological tests). The results of highly sensitive diagnostic techniques with unknown specificity should be interpreted with caution. The organism identified by a new method may represent a real pathogen that was unrecognized by conventional diagnostic methods or contamination during specimen sampling, transportation, or processing. For accurate interpretation, additional studies are needed, which would evaluate the long-term outcome (usually >2 years) with or without antimicrobial treatment. It is expected that new rapid, accurate, and fully automatic diagnostic tests will be developed soon.

Propionibacterium acnes, an emerging pathogen: from acne to implant-infections, from phylotype to resistance.

Propionibacterium acnes colonizes the lipid-rich sebaceous glands of the skin. This preferential anaerobic bacterium is easily identified if cultures are prolonged. It is involved in the inflammation process of acne, but until recently, it was neglected in other clinical presentations. Despite a reported low virulence, the new genomic, transcriptomic, and phylogenetic studies have allowed better understanding of this pathogens importance that causes many chronic and recurrent infections, including orthopedic and cardiac prosthetic, and breast or eye implant-infections. These infections, facilitated by the ability of P. acnes to produce a biofilm, require using anti-biofilm active antibiotics such as rifampicin. The antibiogram of P. acnes is not systematically performed in microbiology laboratories because of its susceptibility to a wide range of antibiotics. However, in the last 10 years, the rate of antibiotic-resistant bacteria has increased, especially for macrolides and tetracyclines. Recently, rpoB gene mutations conferring resistance to rifampicin have been also reported. Thus in case of a biofilm growth mode, the therapeutic strategy should be discussed, according to the resistance phylotype and phenotype so as to optimize the treatment of these severe infections.

Evaluation of 16S rRNA Gene PCR Sensitivity and Specificity for Diagnosis of Prosthetic Joint Infection: a Prospective Multicenter Cross-Sectional Study

ABSTRACT There is no standard method for the diagnosis of prosthetic joint infection (PJI). The contribution of 16S rRNA gene PCR sequencing on a routine basis remains to be defined. We performed a prospective multicenter study to assess the contributions of 16S rRNA gene assays in PJI diagnosis. Over a 2-year period, all patients suspected to have PJIs and a few uninfected patients undergoing primary arthroplasty (control group) were included. Five perioperative samples per patient were collected for culture and 16S rRNA gene PCR sequencing and one for histological examination. Three multicenter quality control assays were performed with both DNA extracts and crushed samples. The diagnosis of PJI was based on clinical, bacteriological, and histological criteria, according to Infectious Diseases Society of America guidelines. A molecular diagnosis was modeled on the bacteriological criterion (≥1 positive sample for strict pathogens and ≥2 for commensal skin flora). Molecular data were analyzed according to the diagnosis of PJI. Between December 2010 and March 2012, 264 suspected cases of PJI and 35 control cases were included. PJI was confirmed in 215/264 suspected cases, 192 (89%) with a bacteriological criterion. The PJIs were monomicrobial (163 cases [85%]; staphylococci, n = 108; streptococci, n = 22; Gram-negative bacilli, n = 16; anaerobes, n = 13; others, n = 4) or polymicrobial (29 cases [15%]). The molecular diagnosis was positive in 151/215 confirmed cases of PJI (143 cases with bacteriological PJI documentation and 8 treated cases without bacteriological documentation) and in 2/49 cases without confirmed PJI (sensitivity, 73.3%; specificity, 95.5%). The 16S rRNA gene PCR assay showed a lack of sensitivity in the diagnosis of PJI on a multicenter routine basis.

Eradication of methicillin-resistant Staphylococcus aureus in a neonatal intensive care unit: which measures for which success?

BACKGROUND Various strategies for controlling methicillin-resistant Staphylococcus aureus (MRSA) outbreaks in neonatal intensive care units (NICUs) have been tried, with varying levels of success. We report a MRSA outbreak occurring between April 2004 and August 2007 in a 24-bed NICU in a large university hospital. We describe the difficulties involved in implementing measures to control the MRSA outbreak and the possible contribution of each measure. METHODS Cases were defined as neonates with MRSA obtained from either clinical cultures or surveillance cultures (from the anterior nares). Systematic screening of neonates for colonization was performed only between February and December 2005. Successive control strategies included barrier precaution and isolation in individual rooms, mupirocine ointment for neonates and health care workers, cohort isolation, hand hygiene observation, and staff training. RESULTS During the routine surveillance culture period (February to December 2005; 48 weeks), 46 neonates were found to be positive for MRSA and were treated with mupirocin. After December 2005, the outbreak was controlled, but the ongoing spread was not eradicated; 9 sporadic MRSA cases were detected by clinical culture up to August 2007. CONCLUSION The widespread use of mupirocine in staff and patients did not control the outbreak and is not recommended. The later control appeared to coincide with increased hand hygiene audits and training for staff, along with appropriate cohort isolation of neonates and cohort nursing.

Retrospective analysis of the risk factors and pathogens associated with early-onset ventilator-associated pneumonia in surgical-ICU head-trauma patients.

Background Early-onset ventilator associated pneumonia (EOVAP) are frequent in head-trauma patients, but specific risk factors are poorly studied in this population. Methods We conducted a retrospective cohort study in a surgical intensive care unit. Consecutive severe head-trauma patients admitted from January 2000 to December 2002 were studied. Microorganisms, and risks factors for EOVAP were analyzed. Results During the 3-year period, 161 patients were studied; 21.1% of them developed an EOVAP. On univariate analysis 6 variables were associated with EOVAP: early enteral feeding, barbiturate use, immunosuppression, mean Simplified Acute Physiology Score 2, acute respiratory distress syndrome, and initial neurosurgery procedures. On multivariate analysis, enteral feeding >2000 Kcal before day 5 [odds ratio (OR): 0.33, 95% confidence interval (CI): 0.21-0.85] and initial neurosurgical procedure (OR: 0.36, 95% CI: 0.15-0.89) remained protective factors for EOVAP, whereas immunosuppression (OR: 7.15, 95% CI: 1.66-30.73) and barbiturate use (OR: 2.68, 95% CI: 1.06-6.80) remained risk factors for EOVAP. EOVAP was also significantly associated with a longer duration of mechanical ventilation (14.0 vs. 11.0 d, P=0.024), and a longer sedation duration (8.3 vs. 5.8 d P=0.005). Methicillin-susceptible Staphylococcus aureus was the most common pathogen involved in EOVAP (46%). Conclusions We demonstrate for the first time that early enteral feeding is a protective factor for EOVAP, and this result could have clinical implications for the prevention of EOVAP after traumatic brain injury. This study also confirms that barbiturate use is an important risk factor of EOVAP whereas Methicillin-susceptible S. aureus was found to be the main pathogen involved in EOVAP.

Significance of Propionibacterium acnes-positive samples in spinal instrumentation.

Study Design. A retrospective study about Propionibacterium acnes infections after Cotrel–Dubousset (CD) instrumentation. Objectives. To analyze the significance of P. acnes-positive deep samples after CD. Summary of Background Data. The diagnosis of spinal infections to P. acnes after CD is difficult. Methods. Patients with revision surgery and at least 1 P. acnes-positive deep sample, between 2000 and 2006 were included. Group A had 1 revision surgery and group B had 2 successive revision surgeries, with P. acnes-positive deep samples. Group A was divided into 2 subgroups according to the peroperative macroscopic aspect, subgroup A1 with septic tissues, subgroup A2 without septic tissues. The biologic characteristics of the patients and the surgical and medical treatments were assessed. Results. Sixty-eight patients were included, 60 in group A (A1 = 33, A2 = 27) and 8 in group B. Group A: 26 patients had 1 or 2 P. acnes-positive samples and 34 had at least 3 P. acnes-positive samples. Histology showed chronic inflammatory changes. C-reactive protein value median rate was 42 (A1) and 5 mg/L (A2). Twenty-two patients had a complete implant removal (14 with antibiotics, A1 = 12, A2 = 2). Nine patients had a total implant replacement (7 with antibiotics). Twenty-two patients had a partial implant removal (17 with antibiotics, A1 = 5, A2 = 12). Seven A1 patients had an irrigation and debridement (6 with antibiotics). The evolution was favorable for 28 patients. Seven patients had a documented sepsis. Group B: during the first revision, 8 patients had a partial implant removal (2 with antibiotics); during the second revision, all patients received antibiotics 4 of whom had a total implant removal. The long-term evolution was favorable for 6 patients. Conclusion. P. acnes infection of spinal instrumentation is difficult to diagnose. Results of at least 4 deep sample cultures, histology, and C-reactive protein values must be compared to the peroperative macroscopic aspect.

Orthopaedic-implant infections by Escherichia coli: molecular and phenotypic analysis of the causative strains.

OBJECTIVES Little is known about Escherichia coli Orthopaedic Implant Infections (OII) pathogenesis. Thus, we compared 30 clinical strains isolated in this context with 30 clinical strains of faecal origin, in order to identify phenotypic and genetic features related to E. coli OII. METHODS Phylogenetic analysis and detection of 19 virulence genes were performed by PCR. Ability to form biofilm was studied using the crystal violet reference method and the innovative BioFilm Ring Test(®). RESULTS Most of the OII isolates (56.7%) belonged to the virulence-associated phylogenetic group B2, but did not present a specific set of virulence factors. S fimbriae was the only adhesin significantly associated with OII isolates. Isolates varied greatly in their ability to form biofilm but OII isolates did not produce significantly more biofilm in vitro than isolates of faecal origin, whatever the method used. CONCLUSIONS Neither a specific pathogenic signature nor an increased ability to form biofilm in vitro was detected in E. coli strains isolated from OII. Nevertheless, genetic properties of these isolates could provide a clue to their origin. Hence, we found that virulence factors of uropathogenic strains and urological disorders were frequently detected among our OII cohort.

−11 Mutation in the ampC Promoter Increasing Resistance to β-Lactams in a Clinical Escherichia coli Strain

ABSTRACT A mutation was discovered in the Pribnow box of the ampC promoter in a clinical Escherichia coli strain. This −11 C-to-T transition created a perfect homology with the −10 consensus sequence. The new promoter was cloned upstream of the cat gene of pKK232-8 and induced a sixfold increase in promoter strength.