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Although carbapenemase-producing Enterobacteriaceae (CPE) have become a serious public health issue, their detection remains challenging. The aim of this study was to implement a test based on imipenem hydrolysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF MS), using 65 strains producing or not a carbapenemase. Then, we compared its performance to that of the Rapidec Carba NP test using 20 additional strains. The MS-based test effectively discriminated between CPE and other non-carbapenem-susceptible strains compared to the Rapidec Carba NP test (sensitivity 100% and 92%, specificity 94% and 92%, respectively). The MS-based test gave less difficulty in interpretation than the colorimetric Rapidec Carba NP test. MALDI-ToF gave a result in less than one hour and limited the use of expensive molecular assays. In conclusion, the hydrolysis test based on MALDI-ToF MS can detect clinically relevant CPE isolates in routine practice. This technology, also described to screen for carbapenem resistance in Pseudomonas aeruginosa and Acinetobacter baumannii complex strains, also seems to be interesting in routine practice for these pathogens.

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Overexpression of efflux pumps extruding antibiotics currently used for the treatment of Acinetobacter baumannii infections has been described as an important mechanism causing antibiotic resistance. The first aim of this work was to phenotypically evaluate the overexpression of efflux pumps on a collection of 124 ciprofloxacin resistant A. baumannii strains. An overexpression of genes encoding one or more efflux pumps was obtained for 19 out of the 34 strains with a positive phenotypic efflux (56%). The most frequent genes overexpressed were those belonging to the RND family, with adeJ being the most prevalent (50%). Interestingly, efflux pump genes coding for MATE and MFS families were also overexpressed quite frequently: abeM (32%) and abaQ (26%). The second aim was to synthesize 1-(1-NaphthylMethyl)-Piperazine analogs as potential new efflux pump inhibitors and biologically evaluate them against strains with a positive phenotypic efflux. Quinoline and pyridine analogs were found to be more effective than their parent compound 1-(1-NaphthylMethyl)-Piperazine. Stereochemistry also played an important part in the inhibitory activity as quinoline derivative ( R )-3a was identified as being the most effective and less cytotoxic. Its inhibitory activity was also correlated to the number of efflux pumps expressed by a strain. The results obtained in this work suggest that quinoline analogs of 1-(1-NaphthylMethyl)-Piperazine are promising leads in the development of new anti- Acinetobacter baumannii therapeutic alternatives, in combination with antibiotics for which an efflux-mediated resistance is suspected.

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Intra-abdominal infections (IAIs) are one of the most common type of infections in patients with sepsis and an important cause of death in intensive care units. Early detection and treatment are necessary to reduce patient complications and improve outcomes. The Unyvero IAI Application (Curetis GmbH) is the first automated assay to rapidly and simultaneously identify a large panel of bacteria, fungi, toxins, and antibiotic resistance markers directly from IAI-related samples. The assay was evaluated in four European clinical laboratories in comparison to routine microbiological practices. A total of 300 clinical samples were tested with an overall sensitivity of 89.3% and specificity of 99.5%, while time to results was reduced by an average of about 17 h compared to identification (ID) results and 41 h compared to full antibiotic susceptibility testing (AST) results. The Unyvero IAI was able to detect additional microorganisms compared with culture, in particular anaerobes, with most detections confirmed by sequencing. The most frequent resistance markers detected were mecA/mecC (n = 25), aacA4 (n = 20), and bla (n = 17) and carbapenemase genes were identified in nine specimens. Further studies are now required to determine the clinical impact of this new rapid test which could play a role in the successful treatment of IAI.

Background Carbapenemase‐producing Enterobacteriaceae (CPE) have become a major public health issue. The objective of the present study was to prospectively assess the analytical performance of a CPE detection algorithm based on phenotypic tests (the screening test) and MALDI‐ToF hydrolysis (the confirmatory test). Methods Over a 6‐month period and based on a disk diffusion method, 74 carbapenem‐resistant strains were included in this study. Results Of the collected isolates, 54 turned out to be negative after phenotypic tests. Hence, 20 strains (including all of the CPEs) were checked with the confirmation test. Seven strains were positive. After molecular biology assessments in a reference center, three of the seven were found to be false positives. The algorithm had a negative predictive value and a sensitivity of 100%, a specificity of 77%, and a positive predictive value of 20%. Conclusion The algorithm has a 24‐hour turnaround time and helps to avoid using expensive molecular biology tests; we consider that it can be used on a routine basis for screening clinical strains.

This systematic review aimed to sum up the evidence gathered by molecular biology methods on the transmission of bacterial clones from sinks/sink drains environmental sources to intensive care unit (ICU) patients. Forty-five reports meeting inclusion/exclusion criteria were identified. Most were retrospective cohort studies on Gram negative multidrug resistant bacteria, with P. aeruginosa and S. marcescens being the most frequent species (26.7% and 17.8% of the studies, respectively). The reports using pulse field gel electrophoresis were the most numerous (44.4%) and found a common clone between clinical and sink/sink drains isolates in 80% of the cases. Over the last 5 years, the use of whole genome sequencing became more frequent and linked sink/sink drains isolates to clinical ones in 50% of the cases. Precise positivity timelines mostly pointed towards a patient-to-sink/sink drain transmission while only 8 reports provided back up for the sink/sink drain-to-patient. To better appraise the role of sinks/sink drains as a reservoir for nosocomial acquisition of bacteria in ICU, future reports should strive to give a precise timeline for the retrieval of isolates as well as the cut-off criteria used to assign isolates to a given clone (information lacking in 66.7% and 42.2% of the studies, respectively).