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      Rapid Carbapenemase Detection With Xpert Carba-R V2 Directly On Positive Blood Vials

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          Abstract

          The rapid detection of carbapenemase allows implementation of infection control measures and adaptation of antibiotic therapy. We evaluated the performances of the Xpert Carba-R V2 ® assay for the direct detection and identification of carbapenemase on positive blood cultures. We focused our evaluation on its detection capacity and on the risks of interference due to the patient’s blood. Isolates of several variants of OXA-48-like (n=10), KPC (n=10), NDM (n=11), VIM (n=7), IMP-1 (n=1) carbapenemases and 14 non carbapenemase-producing Enterobacteriaceae were tested. For each isolate (n=53), an aerobic vial was seeded, and incubated in Bactec Fx (Becton Dickinson ®) automate. When positive, the Xpert ® Carba-R-V2 assay was assessed for carbapenemase detection using 40 µl aliquot. Reproducibility tests were performed on a subset of 23 isolates using aerobic and anaerobic vials. Longer incubation time was also evaluated on 6 isolates. A complementary prospective study in real-time testing of patient-derived clinical samples on 20 additional positive blood vials with Gram negative bacilli on direct examination was performed. Perfect sensitivity and specificity (100%) were observed regardless of the carbapenemase type, the blood vials used and the time of incubation. Xpert ® Carba-R-V2 assay is suitable for the rapid detection of the main carbapenemase genes directly on positive blood vials. Its performances and rapid time analysis allow its use in routine to guide therapeutic choices and to implement infection control measures.

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          Treatment of Infections Caused by Extended-Spectrum-Beta-Lactamase-, AmpC-, and Carbapenemase-Producing Enterobacteriaceae

          Therapy of invasive infections due to multidrug-resistant Enterobacteriaceae (MDR-E) is challenging, and some of the few active drugs are not available in many countries. For extended-spectrum β-lactamase and AmpC producers, carbapenems are the drugs of choice, but alternatives are needed because the rate of carbapenem resistance is rising. Potential active drugs include classic and newer β-lactam–β-lactamase inhibitor combinations, cephamycins, temocillin, aminoglycosides, tigecycline, fosfomycin, and, rarely, fluoroquinolones or trimethoprim-sulfamethoxazole. These drugs might be considered in some specific situations. AmpC producers are resistant to cephamycins, but cefepime is an option. In the case of carbapenemase-producing Enterobacteriaceae (CPE), only some “second-line” drugs, such as polymyxins, tigecycline, aminoglycosides, and fosfomycin, may be active; double carbapenems can also be considered in specific situations. Combination therapy is associated with better outcomes for high-risk patients, such as those in septic shock or with pneumonia. Ceftazidime-avibactam was recently approved and is active against KPC and OXA-48 producers; the available experience is scarce but promising, although development of resistance is a concern. New drugs active against some CPE isolates are in different stages of development, including meropenem-vaborbactam, imipenem-relebactam, plazomicin, cefiderocol, eravacycline, and aztreonam-avibactam. Overall, therapy of MDR-E infection must be individualized according to the susceptibility profile, type, and severity of infection and the features of the patient.
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            Rapid Detection of Carbapenemase-producing Enterobacteriaceae

            To rapidly identify carbapenemase producers in Enterobacteriaceae, we developed the Carba NP test. The test uses isolated bacterial colonies and is based on in vitro hydrolysis of a carbapenem, imipenem. It was 100% sensitive and specific compared with molecular-based techniques. This rapid (<2 hours), inexpensive technique may be implemented in any laboratory.
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              The clinical importance of microbiological findings in the diagnosis and management of bloodstream infections.

              Bloodstream infections are associated with high morbidity and mortality. Accurate identification of blood isolates to the species level and identification of the source of infection and/or the portal of entry are crucial for optimal management of these infections. These investigations-in addition to clinical findings and laboratory and imaging studies-are central to informing and directing efficient and effective diagnostic examinations and to choosing the optimal antimicrobial regimen. Four case studies that demonstrate the importance of identifying the causative agents and the source of infection are discussed to illustrate the central importance of microbiological findings in the diagnosis of bacteremia and bloodstream infections associated with infections at other sites.
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                Author and article information

                Journal
                Infect Drug Resist
                Infect Drug Resist
                IDR
                idr
                Infection and Drug Resistance
                Dove
                1178-6973
                23 October 2019
                2019
                : 12
                : 3311-3316
                Affiliations
                [1 ]IAME, UMR 1137, INSERM, Université Paris Diderot, Sorbonne Paris Cité, AP-HP, Service de Microbiologie, Hôpital Robert-Debré, AP-HP , Paris, France
                [2 ]Service de Microbiologie, Centre National de Référence associé Escherichia coli, Hôpital Robert-Debré, AP-HP , Paris, France
                [3 ]Service de Microbiologie, Hôpitaux Universitaires de Paris Seine Denis (HUPSSD), site Avicenne, AP-HP , Bobigny, France
                [4 ]EA7361, Université Paris-Sud, Université Paris-Saclay, LabEx Lermit, Service de Bactériologie-Hygiène, Hôpital Bicêtre, AP-HP , Le Kremlin-Bicêtre, France
                [5 ]Centre National de Référence associé de la résistance aux antibiotiques: Entérobactéries productrices de carbapénémases , Le Kremlin-Bicêtre, France
                [6 ]Evolution et Ecologie de la résistance aux antibiotiques, Institut Pasteur - APHP -Université Paris Sud , Paris, France
                Author notes
                Correspondence: André Birgy Service de Microbiologie, Hôpital Robert Debré , 48 Boulevard SerrurierParisF75019, FranceTel +33140034060Fax +33140032450 Email andre.birgy@aphp.fr
                Author information
                http://orcid.org/0000-0002-8800-270X
                http://orcid.org/0000-0001-6596-7384
                Article
                204436
                10.2147/IDR.S204436
                6815938
                f39df818-91aa-438e-aff9-7fcdf69a710e
                © 2019 Cointe et al.

                This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License ( http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms ( https://www.dovepress.com/terms.php).

                History
                : 08 February 2019
                : 05 April 2019
                Page count
                Tables: 1, References: 17, Pages: 6
                Categories
                Short Report

                Infectious disease & Microbiology
                carbapenemase,rapid detection,positive blood culture,genexpert,antibiotic therapy,infection control measures

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