Stéphane Paul Emonet

Atypical Infections in Tsunami Survivors

After a tsunami hit Asia in December 2004, 2 survivors had severe infections due to multidrug-resistant and atypical bacteria and rare fungi weeks afterwards. Treating these infections is challenging from a clinical and microbiologic point of view.

Molecular diagnosis of Kingella kingae osteoarticular infections by specific real-time PCR assay.

Kingella kingae is an emerging pathogen that is recognized as a causative agent of septic arthritis and osteomyelitis, primarily in infants and children. The bacterium is best detected by rapid inoculation in blood culture systems or by real-time PCR assays. Pathogenesis of the agent was linked recently to the production of a potent cytotoxin, known as RTX, which is toxic to a variety of human cell types. The locus encoding the RTX toxin is thought to be a putative virulence factor, and is, apparently, essential for inducing cytotoxic effects on respiratory epithelial, synovial and macrophage-like cells. Herein, we describe a novel real-time PCR assay that targets the RTX toxin gene and illustrate its use in two clinical cases. The assay exhibited a sensitivity of 30 c.f.u., which is 10-fold more sensitive than a previously published semi-nested broad-range 16S rRNA gene PCR, and showed no cross-reactivity with several related species and common osteoarticular pathogens.

Evaluation of Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Rapid Identification of Beta-Hemolytic Streptococci

ABSTRACT This study was undertaken to evaluate matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for the rapid identification of beta-hemolytic streptococci. We compared Bruker Biotyper 2.0 with Vitek2 coupled to the agglutination test. MALDI-TOF MS analysis of 386 beta-hemolytic streptococcal isolates yielded high-confidence identification to the species level for all 386 isolates. The Vitek2 gave high-confidence identification to the species level for 88% of Streptococcus agalactiae isolates (n = 269/306), 92% of Streptococcus pyogenes isolates (n = 48/52), and 39% of isolates of Streptococcus dysgalactiae serogroups C and G (n = 11/28).

Low incidence of haematogenous seeding to total hip and knee prostheses in patients with remote infections

OBJECTIVES The exposure of joint prostheses to remote infections is unknown. We wanted to estimate (a) the exposure of arthroplasty patients to severe remote infections, and (b) the incidence of arthroplasty infections associated with remote infections. METHODS Prospective cohort study of all elective hip and knee arthroplasties performed between March 1996 and September 2008, with retrospective documentation of remote infections in hospitalized patients. RESULTS A total of 6101 elective total joint arthroplasties, consisting of 4002 hip replacements (66%) and 2099 knee replacements (34%), were included. The mean follow-up was 70 months. During the study period, the cohort patients experienced 553 remote infections after a median delay of 33 months post-arthroplasty. There were 71 prosthetic infections detected, 7 (total incidence 7/6101, 0.1%) of which were secondary to a remote infection. The ratio of infections associated with remote infections to potential exposure was 1:79. Among hip arthroplasty patients the incidence rate was 1.4 infections associated with remote infections per 10,000 patient-years of follow-up. Infections associated with remote infections occurred later than surgical site infections, (46 months vs. 19 months post-surgery, respectively; mean difference 27 months, 95% CI 8-45 months). CONCLUSIONS Arthroplasty infections associated with remote infections were rare, and occurred like their potential exposure mostly more than 24 months post-arthroplasty.

Evaluation of eight cases of confirmed Bordetella bronchiseptica infection and colonization over a 15-year period

We describe eight human cases of Bordetella bronchiseptica infection and colonization over a 15-year period. Amongst the eight patients, seven had significant underlying disease. Cat exposure was documented in three cases. Symptoms ranged from asymptomatic carriage to severe pneumonia. We could not identify a homogeneous pattern of clinical disease among symptomatic patients. Although B. bronchiseptica infection remains a rare clinical condition among humans, it should be considered as potentially pathogenic when found in airways of immunocompromised patients.

Bordetella holmesii: an under-recognised Bordetella species

Bordetella holmesii, first described in 1995, is believed to cause both invasive infections (bacteraemia, meningitis, endocarditis, pericarditis, pneumonia, and arthritis) and pertussis-like symptoms. Infection with B holmesii is frequently misidentified as being with B pertussis, the cause of whooping cough, because routine diagnostic tests for pertussis are not species-specific. In this Review, we summarise knowledge about B holmesii diagnosis and treatment, and assess research needs. Although no fatal cases of B holmesii have been reported, associated invasive infections can cause substantial morbidities, even in previously healthy individuals. Antimicrobial treatment can be problematic because B holmesiis susceptibility to macrolides (used empirically to treat B pertussis) and third-generation cephalosporins (often used to treat invasive infections) is lower than would be expected. B holmesiis adaptation to human beings is continuing, and virulence might increase, causing the need for better diagnostic assays and epidemiological surveillance.

Development and validation of a modified broad-range 16S rDNA PCR for diagnostic purposes in clinical microbiology

Broad-range PCR followed by sequencing identifies bacterial pathogens, even in challenging settings such as patients receiving antibiotics or infected with fastidious or non-cultivable organisms. The major problem with broad-range PCR is the risk of sample contamination. Risk is present at every step of the procedure, starting from sample collection. Contaminating bacterial DNA may be present not only in laboratory reagents but also at the surface of plastic consumables and containers used for specimen drawing and transport to the diagnostic laboratory. Contaminating DNA is amplified efficiently, leading to false-positive results. Thus, high specificity depends on eliminating such spurious targets, an awkward problem given the abundance of such targets and a highly sensitive method that detects very small numbers of molecules. Several investigators have reported strategies for eliminating the amplification of contaminating DNA sequences. So far, none of these methods has been entirely effective and reproducible. Here we describe a method that uses Exonuclease III (ExoIII) to disable contaminating sequences from acting as templates, while maintaining the high sensitivity of PCR for pathogen DNA. We use this assay in 144 clinical specimens from normally sterile sites, identifying pathogens from 24 (17%). Conventional methods identified pathogens in only four of these specimens, all of which were positive for the same pathogen by PCR. Compared with conventional methods, broad-range PCR with ExoIII pre-treatment of reagents substantially improves the diagnostic yield of bacterial pathogen identification from normally sterile sites.

Randomized non-inferiority trial to compare trimethoprim/sulfamethoxazole plus rifampicin versus linezolid for the treatment of MRSA infection

OBJECTIVES The therapeutic arsenal for MRSA infections is limited. The aim of this study was to assess the non-inferiority of a combination of trimethoprim/sulfamethoxazole plus rifampicin versus linezolid alone for the treatment of MRSA infection. METHODS We conducted a randomized, open-label, single-centre, non-inferiority trial comparing trimethoprim/sulfamethoxazole (160 mg/800 mg three times daily) plus rifampicin (600 mg once a day) versus linezolid (600 mg twice a day) alone in adult patients with various types of MRSA infection. Patients were allocated 1:1 to either regimen. The primary outcome was clinical cure at 6 weeks after the end of treatment (non-inferiority margin 20%) assessed by both ITT and PP analyses. Secondary outcomes included the microbiologically documented persistence of MRSA in clinical cultures, mortality and adverse events. The study protocol has been registered with ClinicalTrials.gov (NCT00711854). RESULTS Overall, 150 patients were randomized to one of the two treatment arms between January 2009 and December 2013 and were included in the ITT analysis. Of these 56/75 (74.7%) in the linezolid group and 59/75 (78.7%) in the trimethoprim/sulfamethoxazole and rifampicin group experienced clinical success (risk difference 4%, 95% CI -9.7% to 17.6%). The results were confirmed by the PP analysis, with 54/66 (81.8%) cured patients in the linezolid group versus 52/59 (88.1%) in the trimethoprim/sulfamethoxazole and rifampicin group (risk difference 6.3%, 95% CI -6.8% to 19.2%). There were no statistically significant differences between the two groups in any of the secondary outcomes, including microbiologically documented failure. Four adverse drug reactions attributed to the study medication occurred in the linezolid group versus nine in the trimethoprim/sulfamethoxazole and rifampicin group. CONCLUSIONS Compared with linezolid, trimethoprim/sulfamethoxazole and rifampicin seems to be non-inferior in the treatment of MRSA infection.

The role of anaerobes in diabetic foot infections.

Diabetic foot infections (DFI) are a common cause of morbidity and, on occasion, even mortality. Infection can be either mono- or polymicrobial, with a wide variety of potential pathogens. Anaerobes may be involved, particularly in wounds that are deeper or more chronic, and are more frequently identified when using modern molecular techniques, such as 16s PCR and pyrosequencing. It remains unclear whether the presence of anaerobes in DFI leads to more severe manifestations, or if these organisms are largely colonizers associated with the presence of greater degrees of tissue ischemia and necrosis. Commonly used empiric antibiotic therapy for diabetic foot infections is generally broad-spectrum and usually has activity against the most frequently identified anaerobes, such as Peptostreptococcus and Bacteroides species. Adequate surgical debridement and, when needed, foot revascularization may be at least as important as the choice of antibiotic to achieve a successful treatment outcome.

Building a transnational biosurveillance network using semantic web technologies: requirements, design, and preliminary evaluation.

Background Antimicrobial resistance has reached globally alarming levels and is becoming a major public health threat. Lack of efficacious antimicrobial resistance surveillance systems was identified as one of the causes of increasing resistance, due to the lag time between new resistances and alerts to care providers. Several initiatives to track drug resistance evolution have been developed. However, no effective real-time and source-independent antimicrobial resistance monitoring system is available publicly. Objective To design and implement an architecture that can provide real-time and source-independent antimicrobial resistance monitoring to support transnational resistance surveillance. In particular, we investigated the use of a Semantic Web-based model to foster integration and interoperability of interinstitutional and cross-border microbiology laboratory databases. Methods Following the agile software development methodology, we derived the main requirements needed for effective antimicrobial resistance monitoring, from which we proposed a decentralized monitoring architecture based on the Semantic Web stack. The architecture uses an ontology-driven approach to promote the integration of a network of sentinel hospitals or laboratories. Local databases are wrapped into semantic data repositories that automatically expose local computing-formalized laboratory information in the Web. A central source mediator, based on local reasoning, coordinates the access to the semantic end points. On the user side, a user-friendly Web interface provides access and graphical visualization to the integrated views. Results We designed and implemented the online Antimicrobial Resistance Trend Monitoring System (ARTEMIS) in a pilot network of seven European health care institutions sharing 70+ million triples of information about drug resistance and consumption. Evaluation of the computing performance of the mediator demonstrated that, on average, query response time was a few seconds (mean 4.3, SD 0.1×102 seconds). Clinical pertinence assessment showed that resistance trends automatically calculated by ARTEMIS had a strong positive correlation with the European Antimicrobial Resistance Surveillance Network (EARS-Net) (ρ = .86, P < .001) and the Sentinel Surveillance of Antibiotic Resistance in Switzerland (SEARCH) (ρ = .84, P < .001) systems. Furthermore, mean resistance rates extracted by ARTEMIS were not significantly different from those of either EARS-Net (∆ = ±0.130; 95% confidence interval –0 to 0.030; P < .001) or SEARCH (∆ = ±0.042; 95% confidence interval –0.004 to 0.028; P = .004). Conclusions We introduce a distributed monitoring architecture that can be used to build transnational antimicrobial resistance surveillance networks. Results indicated that the Semantic Web-based approach provided an efficient and reliable solution for development of eHealth architectures that enable online antimicrobial resistance monitoring from heterogeneous data sources. In future, we expect that more health care institutions can join the ARTEMIS network so that it can provide a large European and wider biosurveillance network that can be used to detect emerging bacterial resistance in a multinational context and support public health actions.