Comparison of Xpert MRSA/SA Nasal and MRSA/SA ELITe MGB assays for detection of the mecA gene with susceptibility testing methods for determination of methicillin resistance in Staphylococcus aureus isolates.

In a series of 82 Staphylococcus strains isolated from culture, 100% were identified as Staphylococcus aureus by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS); 99.9% (77/82) of them were resistant to benzylpenicillin, oxacillin, and cefoxitin, and 6.1% (5/82) were susceptible to methicillin. Xpert MRSA/SA assay results were concordant with the phenotypic results in 76.8% (63/82) of cases and discordant in 23.2% (19/82) of cases. The MRSA/SA ELITe MGB kit results were concordant with phenotypic results in 100% of the cases. When comparing the Xpert MRSA/SA assay results with the MRSA/SA ELITe MGB kit results, 78% (64/82) of the cases were concordant, while 22% (18/82) of the cases were discordant. No statistically significant differences were observed between the two techniques. The PCR protocol that was used to validate the results of these two methods gave the following results: 49 were conventional methicillin-resistant S. aureus (MRSA) isolates (mecA positive and mecALGA251 negative), and 25 were phenotypic MRSA isolates (mecA negative and mecALGA251 positive).

Molecular typing and characterization of extended-spectrum TEM, SHV and CTX-M beta-lactamases in clinical isolates of Enterobacter cloacae.

Sixty-one non-repetitive Enterobacter cloacae ESBL producers were collected at the Amiens University Hospital in France. Eight beta-lactam resistance phenotypes (a-h) and three aminoglycoside resistance phenotypes (i-k) were identified among these isolates, and 32 different pulsotypes were observed. Of these 61 isolates, 37 were sequenced and found to harbor beta-lactamases with a pI of 5.9 (TEM-4), 6.5 (TEM-24), 7.8 (SHV-4), 8.2 (SHV-12), 8.4 (CTX-M-1) and 8.0 (CTX-M-9). Four imipenem-resistant ESBL-producing E. cloacae isolates did not express the 38kDa OMP, indicating that this resistance is associated with porin deficiency.

Molecular characterisation and mechanisms of resistance of multidrug-resistant human Salmonella enterica serovar Typhimurium isolated in Amiens (France).

Antimicrobial resistance patterns of Salmonella enterica serovar Typhimurium isolates obtained during the study period were examined. The molecular epidemiology and the mechanisms of resistance to ampicillin, chloramphenicol and tetracycline were investigated. Resistance to ampicillin increased from 59% between 1996 and 1999 to 62.5% in 2000 and to 66.6% in 2001. Of 51 S. Typhimurium isolates studied, 100% were resistant to ampicillin (minimum inhibitory concentration (MIC)>256 mg/L) and sulphonamide (MIC range, 128 to >256 mg/L). Ninety-eight percent of isolates were resistant to streptomycin (MIC range, 48-256 mg/L), 92.2% to tetracycline (MIC range, 32 to >256 mg/L), 88.2% to chloramphenicol (MIC>256 mg/L), 21.5% to sulphamethoxazole/trimethoprim (MIC>32 mg/L), 5.8% to amoxicillin/clavulanic acid (MIC, 32 mg/L) and 1.9% to cefalothin (MIC, 64mg/L). Six resistance phenotypes were found (a-f), with phenotypes a (47%) and b (27.5%) being predominant. Twenty-five (49%) of 51 isolates produced a single beta-lactamase, among which 48% produced PSE-1, 44% produced TEM-1 and 8% produced OXA-1. Among 26 of the 51 isolates, 10 produced PSE-1+OXA-1, 7 produced TEM-1+PSE-1+OXA-1, 6 produced TEM-1+PSE-1, and 3 produced TEM-1+OXA-1. Forty-eight (94.1%) of the 51 isolates had the plasmid-mediated resistance gene flo(ST) to chloramphenicol and tetracycline. Combining enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) and pulsed-field gel electrophoresis (PFGE), 16 distinct patterns were identified, among which patterns IA (35.3%) and IF (27.4%) were considered as epidemic patterns. The dendrogram obtained from S. Typhimurium pulsotypes allowed five clones (S1-S5) to be identified, with two prevalent clones comprising 47.8% (S2) and 27.3% (S4) of the isolates.

The usefulness of 99mTc sulfur colloid bone marrow scintigraphy combined with 111In leucocyte scintigraphy in prosthetic joint infection.

AIM: To assess the extent to which bone marrow scintigraphy (BMS) makes the interpretation of leucocyte scintigraphy (LS) easier and improves its diagnostic value. METHODS: Seventy-three 111In LSs, 99mTc hydroxymethylene diphosphonate bone scintigraphies (BSs) and 99mTc sulfur colloid BMSs were performed in 60 patients with suspected infection related to a hip prosthesis or knee prosthesis, either in situ (+group, n = 43) or after removal for septic loosening (-group, n = 30). Bacteriological samples were obtained from all patients. LS was interpreted together with BS (LS-BS) or with BMS (LS-BMS) by three independent readers. RESULTS: The concordance among readers, estimated by the kappa test, was average with LS-BS (kappa/kappam coefficients = 0.58, 0.58 and 0.46, respectively, for the three pairs of readers) and excellent with LS-BMS (kappa/kappam coefficients = 1.00 for the three pairs of readers). With LS-BS, 64/219 interpretations were equivocal whereas only one was equivocal with LS-BMS. Sensitivity, specificity and accuracy of LS-BMS were, respectively, 80%, 94% and 91% in the +group, and 33%, 100% and 93% in the -group. CONCLUSION: We conclude that (1) the interpretation of the results for LS-BMS is very easy, in contrast to LS-BS; (2) the diagnostic value of LS-BMS for detecting infected joint prostheses is good; and (3) additional data are needed to assess the accuracy of LS-BMS when the prosthesis has been removed.

Value of Xpert MRSA/SA blood culture assay on the Gene Xpert® Dx System for rapid detection of Staphylococcus aureus and coagulase-negative staphylococci in patients with staphylococcal bacteremia.

The Xpert MRSA/SA BC assay was examined prospectively in patients with staphylococcal bacteremia including 6 patients with blood culture bottles inoculated with biological fluid (synovial fluid in 4 cases and peritoneal fluid in 2 cases). Among the 56 Staphylococci species isolated, 80.3% were coagulase-negative staphylococci (CoNS) and 19.7% were S. aureus. Methicillin susceptibility test results showed that 77.8% of isolates were methicillin-resistant CoNS (MRCoNS) and 22.2% of isolates were methicillin-susceptible CoNS (MSCoNS). Of 11 S. aureus isolates, 63.7% were methicillin-susceptible S. aureus (MSSA) and 36.3% were methicillin-resistant S. aureus (MRSA). Xpert MRSA/SA BC results showed that genotypic results were in concordance with phenotypic results in 94.6% of cases versus 5.4% of discordant cases. Of these 3 discordant cases, 1 S. aureus isolate had an MRSA phenotype and an SPA(+)mec(+)SCCmec(-) genotype and another S. aureus isolate was phenotypically MSSA and genotypically SPA(+)mec(+)SCCmec(-), and 1 S. epidermidis isolate was phenotypically MSCoNS and genotypically SPA(-)mec(+)SCCmec(-).

First case of human spondylodiscitis due to Shewanella algae.

We present the first case of human spondylodiscitis due to Shewanella algae. Our patient did not have any predisposing factors. The portal of entry was probably a cutaneous lesion on the leg, exposed to seawater. Bacteria were isolated in pure culture from a needle biopsy specimen of the vertebral disk. Automated identification systems identified the organism as Shewanella putrefaciens. However, molecular biology identified it as S. algae. Treatment with ceftriaxone and amikacin, then ciprofloxacin successfully addressed the infection. We also review four published cases of human osteoarticular infections caused by Shewanella spp: two cases of arthritis and two cases of osteomyelitis. Two patients had predisposing factors, and contact with water was found in two cases. The clinical, radiological and biological characteristics of S. algae spondylodiscitis are indistinguishable from those of spondylodiscitis of other causes. A cutaneous lesion with exposure to water is a potential portal of entry. Molecular typing is necessary to obtain a precise bacteriological identification.

Nocardia nova as the causative agent in spondylodiscitis and psoas abscess.

We describe here the first case of Nocardia nova spondylodiscitis accompanied by a psoas abscess due to spreading from pulmonary nocardiosis. Nocardia was cultured from all affected sites. After 1 year of an appropriate antimicrobial therapy and a surgical drainage of the abscess that was required, the patient's clinical condition had improved.