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.

These results suggest that early discontinuation of empirical antibiotics in FUO is safe for afebrile neutropenic patients.

The recognition of carbapenemase-producing Enterobacteriaceae (CPE) isolates is a major laboratory challenge, and their inappropriate or delayed detection may have negative impacts on patient management and on the implementation of infection control measures. We describe here a matrix-assisted laser desorption ionization؊time of flight (MALDI-TOF)-based method to detect carbapenemase activity in Enterobacteriaceae. After a 20-min incubation of the isolate with 0.5 mg/ml imipenem at 37°C, supernatants were analyzed by MALDI-TOF in order to identify peaks corresponding to imipenem (300 Da) and an imipenem metabolite (254 Da). A total of 223 strains, 77 CPE (OXA-48 variants, KPC, NDM, VIM, IMI, IMP, and NMC-A) and 146 non-CPE (cephalosporinases, extended-spectrum ␤-lactamases [ESBLs], and porin defects), were tested and used to calculate a ratio of imipenem hydrolysis: mass spectrometry [MS] ratio ‫؍‬ metabolite/(imipenem ؉ metabolite). An MS ratio cutoff was statistically determined to classify strains as carbapenemase producers (MS ratio of >0.82). We validated this method first by testing 30 of our 223 isolates (15 CPE and 15 non-CPE) 10 times to calculate an intraclass correlation coefficient (ICC of 0.98), showing the excellent repeatability of the method. Second, 43 strains (25 CPE and 18 non-CPE) different from the 223 strains used to calculate the ratio cutoff were used as external controls and blind tested. They yielded sensitivity and specificity of 100%. The total cost per test is <0.10 U.S. dollars (USD). This easy-to-perform assay is time-saving, cost-efficient, and highly reliable and might be used in any routine laboratory, given the availability of mass spectrometry, to detect CPE.

h Although numerous perioperative samples and culture media are required to diagnose prosthetic joint infection (PJI), their exact number and types have not yet been definitely determined with a high level of proof. We conducted a prospective multicenter study to determine the minimal number of samples and culture media required for accurate diagnosis of PJI. Over a 2-year period, consecutive patients with clinical signs suggesting PJI were included, with five perioperative samples per patient. The bacteriological and PJI diagnosis criteria were assessed using a random selection of two, three, or four samples and compared with those obtained using the recommended five samples (references guidelines). The results obtained with two or three culture media were then compared with those obtained with five culture media for both criteria. The times-to-positivity of the different culture media were calculated. PJI was confirmed in 215/264 suspected cases, with a bacteriological criterion in 192 (89%). The PJI was monomicrobial (85%) or polymicrobial (15%). Percentages of agreement of 98.1% and 99.7%, respectively, for the bacteriological criterion and confirmed PJI diagnosis were obtained when four perioperative samples were considered. The highest percentages of agreement were obtained with the association of three culture media, a blood culture bottle, a chocolate agar plate, and Schaedler broth, incubated for 5, 7, and 14 days, respectively. This new procedure leads to significant cost saving. Our prospective multicenter study showed that four samples seeded on three culture media are sufficient for diagnosing PJI. There is still no standard definition of prosthetic joint infection (PJI). Although numerous perioperative samples are needed to diagnose PJI, their optimal number is not yet definitively known. In 1998, Atkins et al., using a mathematical model, demonstrated that five or six perioperative samples, with a cutoff of three or more culture-positive samples, are needed to diagnose PJI with high sensitivity and specificity (1). Recent guidelines still recommend that about five samples should be cultured for optimal diagnosis of PJI (2, 3). Since each tissue specimen is cultured on four to five culture media, a total of 20 to 25 culture media have to be analyzed per patient. The optimal duration of incubation for periprosthetic cultures was extended to 13 or 14 days, especially for Propionibacterium acnes recovery (4, 5). The implementation of 14-day culture incubation in combination with the use of multiple solid and liquid aerobic and anaerobic culture media makes the daily monitoring of PJI by the laboratory very complex, costly, and time-consuming. Moreover, the number of cases of PJI will continue to increase in the coming decades. Forecasts predict that by 2030, the demand for total hip and knee arthroplasties will grow to 4 million in the United States (6). A major economic effect is that device-associated costs are significantly higher than native bone and joint infections, as recently reported (7). Thi...

The epidemiology of carbapenemases worldwide is showing that OXA-48 variants are becoming the predominant carbapenemase type in Enterobacteriaceae in many countries. However, not all OXA-48 variants possess significant activity toward carbapenems (e.g., OXA-163). Two Serratia marcescens isolates with resistance either to carbapenems or to extended-spectrum cephalosporins were successively recovered from the same patient. A genomic comparison using pulsed-field gel electrophoresis and automated Rep-PCR typing identified a 97.8% similarity between the two isolates. Both strains were resistant to penicillins and first-generation cephalosporins. The first isolate was susceptible to expanded-spectrum cephalosporins, was resistant to carbapenems, and had a significant carbapenemase activity (positive Carba NP test) related to the expression of OXA-48. The second isolate was resistant to expanded-spectrum cephalosporins, was susceptible to carbapenems, and did not express a significant imipenemase activity, (negative for the Carba NP test) despite possessing a bla OXA-48 -type gene. Sequencing identified a novel OXA-48-type ␤-lactamase, OXA-405, with a four-amino-acid deletion compared to OXA-48. The bla OXA-405 gene was located on a ca. 46-kb plasmid identical to the prototype IncL/M bla OXA-48 -carrying plasmid except for a ca. 16.4-kb deletion in the tra operon, leading to the suppression of self-conjugation properties. Biochemical analysis showed that OXA-405 has clavulanic acid-inhibited activity toward expanded-spectrum activity without significant imipenemase activity. This is the first identification of a successive switch of catalytic activity in OXA-48-like ␤-lactamases, suggesting their plasticity. Therefore, this report suggests that the first-line screening of carbapenemase producers in Enterobacteriaceae may be based on the biochemical detection of carbapenemase activity in clinical settings.A mbler class D ␤-lactamases (oxacillinases) are widely disseminated among clinically relevant Gram-negative bacteria (1). They exhibit a high degree of diversity of hydrolysis activity ranging from narrow to broad-spectrum hydrolysis activity toward ␤-lactams (1). Among the class D ␤-lactamases, several enzymes hydrolyze carbapenems. Most carbapenemhydrolyzing class D ␤-lactamases (CHDLs) are from Acinetobacter spp. (e.g., OXA-23, OXA-40, OXA-58, OXA-143) (2, 3), whereas OXA-48-type enzymes are identified in Enterobacteriaceae only (4). The OXA-48-derived CHDLs have initially been identified in Turkey (5), first in Klebsiella pneumoniae and then in other enterobacterial species (4). The known OXA-48 variants are currently as follows: (i) OXA-162, identified from K. pneumoniae isolates in Turkey (6); (ii) OXA-163, identified from K. pneumoniae and Enterobacter cloacae isolates in Argentina (7, 8); (iii) OXA-181, identified from a K. pneumoniae isolate in India (9); (iv) OXA-204, identified from K. pneumoniae isolates in patients having a link with North Africa (10); (v) OXA-232, identified in France from a K. pneumoni...

cultures of Salmonella enterica serotype Babelsberg and 6 cultures of serotype Enteritidis, isolated in France from internationally adopted children, were identified at the French National Reference Center for Salmonella. All serotype Babelsberg isolates were related, as determined by pulsed-field gel electrophoresis, and all serotype Enteritidis strains displayed the same phage type. All serotype Enteritidis and seven serotype Babelsberg isolates produced an SHV-12-like extended-spectrum ␤-lactamase as determined by sequencing of PCR products and by isoelectrofocusing. Some serotype Enteritidis isolates exhibited additional antimicrobial resistance (aminoglycosides, tetracycline, chloramphenicol, sulfonamides, and trimethoprim). Our investigation indicated that these Salmonella isolates were certainly acquired in the same orphanage in Bamako, Mali, before the children were adopted by French families. An inappropriate use of ceftriaxone was probably the cause of the emergence of such strains. There is an urgent need to determine the origin of the contamination and to introduce adequate antibiotic protocols into this orphanage to prevent further transmission and dissemination. Screening for infections and follow-up, adapted to the origin of the internationally adopted children, should be recommended.

Six strains of Staphylococcus aureus isolated from cystic fibrosis patients after treatment with azithromycin were cross-resistant to azithromycin and erythromycin. None of the isolates contained erm or msr(A) genes, but they all carried either A2058G/U or A2059G mutations within the rrl genes, with a majority of the rRNA copies bearing the mutation. One strain displayed an additional mutation in the rplV gene, encoding the L22 ribosomal protein

International travellers, transfer of patients and now adoption may contribute to the global emergence of MDR bacteria. Thus, in addition to the usual screening of adopted children for infectious diseases, additional screening for MDR bacteria should be recommended, at least for children coming from countries with a high prevalence of MDR bacteria.