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Patrice Nordmann - One of the best experts on this subject based on the ideXlab platform.
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NitroSpeed-Carba NP Test for Rapid Detection and Differentiation between Different Classes of Carbapenemases in Enterobacterales
Journal of clinical microbiology, 2020Co-Authors: Patrice Nordmann, Mustafa Sadek, Anthony Demord, Laurent PoirelAbstract:A biochemical test (NitroSpeed-Carba NP test) was developed to identify Carbapenemase production in Enterobacterales and to discriminate between the different types of clinically-significant Carbapenemases (Ambler class A, B and D). It is based on two main features, namely the hydrolysis by all s-lactamases including Carbapenemases of the nitrocefin substrate, and the capacity of ertapenem to prevent this hydrolysis for all s-lactamases except Carbapenemases. Specific Carbapenemase inhibitors of class A (avibactam, vaborbactam), class B (dipicolinic acid) and class D (avibactam) were used to inhibit the nitrocefin hydrolysis and to allow the identification of the Carbapenemase types with a turn-around-time of ca. 30 min. The test was evaluated with a collection of 248 clinical enterobacterial isolates including 148 Carbapenemase producers and 100 non-Carbapenemase producers. Its overall sensitivity and specificity were 100% and 97.1%, respectively, including detection of all types of OXA-48-like Carbapenemases. For the detection of the Carbapenemase type including strains that produce double Carbapenemases, the sensitivity was 100%, 97% and 100% for the detection of class A, B, and D, respectively. This easy-to-implement test may contribute to optimize the choice of the s-lactam/s-lactamase inhibitor combinations for treating infection due to Carbapenemase producers.
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rapidec carba np test for rapid detection of Carbapenemase producers
Journal of Clinical Microbiology, 2015Co-Authors: Laurent Poirel, Patrice NordmannAbstract:Performances of the Rapidec Carba NP test (bioMerieux) were evaluated for detection of all types of Carbapenemases in Enterobacteriaceae, Acinetobacter baumannii, and Pseudomonas aeruginosa. In less than 2 h after sample preparation, it showed a sensitivity and specificity of 96%. This ready-to-use test is well adapted to the daily need for detection of Carbapenemase producers in any laboratory worldwide.
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CarbAcineto NP Test for Rapid Detection of Carbapenemase-Producing Acinetobacter spp.
Journal of clinical microbiology, 2014Co-Authors: Laurent Dortet, Laurent Poirel, Caroline Errera, Patrice NordmannAbstract:Multidrug-resistant Acinetobacter baumannii isolates, particularly those that produce Carbapenemases, are increasingly reported worldwide. The biochemically based Carba NP test, extensively validated for the detection of Carbapenemase producers among Enterobacteriaceae and Pseudomonas spp., has been modified to detect Carbapenemase production in Acinetobacter spp. A collection of 151 Carbapenemase-producing and 69 non-Carbapenemase-producing Acinetobacter spp. were tested using the Carba NP test and a modified Carba NP protocol (the CarbAcineto NP test) in this study. The CarbAcineto NP test requires modified lysis conditions and an increased bacterial inoculum compared to those of the original Carba NP test. The Carba NP test detects metallo-β-lactamase producers but failed to detect the production of other Carbapenemase types among Acinetobacter spp. In contrast, the newly designed CarbAcineto NP test, which is rapid and reproducible, detects all types of Carbapenemases with a sensitivity of 94.7% and a specificity of 100%. This cost-effective technique offers a reliable and affordable technique for identifying Carbapenemase production in Acinetobacter spp., which is a marker of multidrug resistance in those species. Its use will facilitate the recognition of these Carbapenemases and prevent their spread.
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Carbapenemase producing enterobacteriaceae overview of a major public health challenge
Medecine Et Maladies Infectieuses, 2014Co-Authors: Patrice NordmannAbstract:Bacterial resistance to antibiotics has become a major source of concern for public health. Enterobacteriaceae are among the most common human pathogens, causing community-acquired as well as hospital-acquired infections. Carbapenem-resistant Enterobacteriaceae have been increasingly reported worldwide since their first identification more than 20 years ago. Three main classes of Carbapenemases have been identified: Ambler class A beta-lactamase (KPC), class B (metallo-enzymes), and class D (OXA-48 type). Klebsiella pneumoniae Carbapenemases (KPC) was first reported in the United States in the late 1990s and since then worldwide, with a marked endemicity in the United States, Greece, and now Italy. Carbapenemase NDM-1 (New Delhi metallo-beta-lactamase-1) is one of the most recently reported metallo-enzymes. It has spread widely in the Indian sub-continent and now worldwide. Carbapenemases of the oxacillinase-48 type (OXA-48) have been identified mostly in Mediterranean and southern European countries with a rapid spread. An early and quick identification of Carbapenemase-producing infected patients, but also of carriers, is mandatory to prevent the spread of these highly resistant pathogens. The early identification of carriers and implementing of cohorting strategies is the only means to prevent nosocomial outbreaks caused by Carbapenemase, with very few, if any, therapeutic options.
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Dissemination of Carbapenemase-producing Enterobacteriaceae in France, 2012
The Journal of antimicrobial chemotherapy, 2013Co-Authors: Laurent Dortet, Gaelle Cuzon, Patrice NordmannAbstract:Objectives Carbapenem-resistant Enterobacteriaceae isolates (n = 1485) were received at the French Associated National Reference Center for Antibiotic Resistance in 2012 and were characterized for their mechanism of resistance to carbapenems. Methods Carbapenemase production was detected using the biochemical-based Carba NP test, based on the detection of in vitro hydrolysis of imipenem. All isolates with a positive Carba NP test result were characterized by PCR and sequencing. Results Carbapenemase production was identified in 23.1% of the isolates. The main Carbapenemase type identified was OXA-48 and derivatives (75.5%). An overseas source was clearly demonstrated for only 27.6% of the isolates. Conclusions OXA-48 and derivatives are now the most prevalent Carbapenemases in France, with a possible spread of these producers in the community.
Laurent Poirel - One of the best experts on this subject based on the ideXlab platform.
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NitroSpeed-Carba NP Test for Rapid Detection and Differentiation between Different Classes of Carbapenemases in Enterobacterales
Journal of clinical microbiology, 2020Co-Authors: Patrice Nordmann, Mustafa Sadek, Anthony Demord, Laurent PoirelAbstract:A biochemical test (NitroSpeed-Carba NP test) was developed to identify Carbapenemase production in Enterobacterales and to discriminate between the different types of clinically-significant Carbapenemases (Ambler class A, B and D). It is based on two main features, namely the hydrolysis by all s-lactamases including Carbapenemases of the nitrocefin substrate, and the capacity of ertapenem to prevent this hydrolysis for all s-lactamases except Carbapenemases. Specific Carbapenemase inhibitors of class A (avibactam, vaborbactam), class B (dipicolinic acid) and class D (avibactam) were used to inhibit the nitrocefin hydrolysis and to allow the identification of the Carbapenemase types with a turn-around-time of ca. 30 min. The test was evaluated with a collection of 248 clinical enterobacterial isolates including 148 Carbapenemase producers and 100 non-Carbapenemase producers. Its overall sensitivity and specificity were 100% and 97.1%, respectively, including detection of all types of OXA-48-like Carbapenemases. For the detection of the Carbapenemase type including strains that produce double Carbapenemases, the sensitivity was 100%, 97% and 100% for the detection of class A, B, and D, respectively. This easy-to-implement test may contribute to optimize the choice of the s-lactam/s-lactamase inhibitor combinations for treating infection due to Carbapenemase producers.
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Carbapenemase-Producing Organisms: A Global Scourge
Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 2017Co-Authors: Robert A. Bonomo, Brandi Limbago, Laurent Poirel, Eileen M. Burd, John Conly, Julie A. Segre, Lars F. WestbladeAbstract:The dramatic increase in the prevalence and clinical impact of infections caused by bacteria producing Carbapenemases is a global health concern. Carbapenemase production is especially problematic when encountered in members of the family Enterobacteriaceae. Due to their ability to readily spread and colonize patients in healthcare environments, preventing the transmission of these organisms is a major public health initiative and coordinated international effort are needed. Central to the treatment and control of Carbapenemase-producing organisms (CPOs) are phenotypic (growth-/biochemical-dependent) and nucleic acid-based Carbapenemase detection tests that identify Carbapenemase activity directly or their associated molecular determinants. Importantly, bacterial isolates harboring Carbapenemases are often resistant to multiple antibiotic classes, resulting in limited therapy options. Emerging agents, novel antibiotic combinations and treatment regimens offer promise for management of these infections. This review highlights our current understanding of CPOs with emphasis on their epidemiology, detection, treatment, and control.
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FRI-2 Carbapenemase-producing Enterobacter cloacae complex in the UK.
The Journal of antimicrobial chemotherapy, 2017Co-Authors: Danièle Meunier, Laurent Poirel, Jacqueline Findlay, Michel Doumith, Daniel Godoy, Claire Perry, Rachel Pike, Firza Gronthoud, Theresa Shryane, William WelfareAbstract:Objectives Detection of rarer Carbapenemases is challenging, as it requires molecular assays with comprehensive coverage or the use of phenotypic methods for the detection of Carbapenemase activity. We describe a new class A Carbapenemase, FRI-2, in an Enterobacter cloacae complex isolate following implementation of an in-house multiplex PCR for the detection of 'rare' class A Carbapenemases. Methods MICs were determined by agar dilution. A carbapenem-resistant E. cloacae complex isolate was tested by PCR for the class A Carbapenemases blaKPC, blaFRI, blaIMI, blaGES and blaSME. Carbapenemase activity was assessed using Carba NP and the carbapenem inactivation method. Whole genome and plasmid analyses of the clinical isolate and the FRI-2 transformant were performed by WGS, respectively. Typing was carried out by PFGE. Results The E. cloacae complex isolate showed resistance to imipenem (MIC = 16 mg/L), meropenem (MIC = 8 mg/L) and ertapenem (MIC = 8 mg/L), but remained susceptible to piperacillin/tazobactam (MIC = 8 mg/L). Carbapenemase activity was confirmed in the isolate by both phenotypic methods. A blaFRI-1-like gene was detected by PCR and analysis of WGS data of the clinical isolate identified an ORF of 885 bp, which showed 97% nucleotide identity with blaFRI-1 and was named blaFRI-2. WGS of the transformant indicated blaFRI-2 was located on a 108 kb IncF/IncR plasmid. The FRI-2-positive E. cloacae complex isolate belonged to a novel ST (ST829). Conclusions The possible circulation of rarer Carbapenemases in clinical settings highlights the role of phenotypic tests to detect Carbapenemase activity when molecular assays are negative for the 'big 5' Carbapenemase families.
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rapidec carba np test for rapid detection of Carbapenemase producers
Journal of Clinical Microbiology, 2015Co-Authors: Laurent Poirel, Patrice NordmannAbstract:Performances of the Rapidec Carba NP test (bioMerieux) were evaluated for detection of all types of Carbapenemases in Enterobacteriaceae, Acinetobacter baumannii, and Pseudomonas aeruginosa. In less than 2 h after sample preparation, it showed a sensitivity and specificity of 96%. This ready-to-use test is well adapted to the daily need for detection of Carbapenemase producers in any laboratory worldwide.
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CarbAcineto NP Test for Rapid Detection of Carbapenemase-Producing Acinetobacter spp.
Journal of clinical microbiology, 2014Co-Authors: Laurent Dortet, Laurent Poirel, Caroline Errera, Patrice NordmannAbstract:Multidrug-resistant Acinetobacter baumannii isolates, particularly those that produce Carbapenemases, are increasingly reported worldwide. The biochemically based Carba NP test, extensively validated for the detection of Carbapenemase producers among Enterobacteriaceae and Pseudomonas spp., has been modified to detect Carbapenemase production in Acinetobacter spp. A collection of 151 Carbapenemase-producing and 69 non-Carbapenemase-producing Acinetobacter spp. were tested using the Carba NP test and a modified Carba NP protocol (the CarbAcineto NP test) in this study. The CarbAcineto NP test requires modified lysis conditions and an increased bacterial inoculum compared to those of the original Carba NP test. The Carba NP test detects metallo-β-lactamase producers but failed to detect the production of other Carbapenemase types among Acinetobacter spp. In contrast, the newly designed CarbAcineto NP test, which is rapid and reproducible, detects all types of Carbapenemases with a sensitivity of 94.7% and a specificity of 100%. This cost-effective technique offers a reliable and affordable technique for identifying Carbapenemase production in Acinetobacter spp., which is a marker of multidrug resistance in those species. Its use will facilitate the recognition of these Carbapenemases and prevent their spread.
Gian Maria Rossolini - One of the best experts on this subject based on the ideXlab platform.
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colistin resistance superimposed to endemic carbapenem resistant klebsiella pneumoniae a rapidly evolving problem in italy november 2013 to april 2014
Eurosurveillance, 2014Co-Authors: M Monaco, M Raffone, Fabio Arena, Aurora Garciafernandez, Simona Pollini, Hajo Grundmann, Annalisa Pantosti, Tommaso Giani, Gian Maria RossoliniAbstract:Consecutive non-replicate clinical isolates (n=191) of carbapenem non-susceptible Enterobacteriaceae were collected from 21 hospital laboratories across Italy from November 2013 to April 2014 as part of the European Survey on Carbapenemase-producing Enterobacteriaceae (EuSCAPE) project. Klebsiella pneumonia Carbapenemase-producing K. pneumoniae (KPC-KP) represented 178 (93%) isolates with 76 (43%) respectively resistant to colistin, a key drug for treating carbapenamase-producing Enterobacteriaceae. KPC-KP colistin-resistant isolates were detected in all participating laboratories. This underscores a concerning evolution of colistin resistance in a setting of high KPC-KP endemicity.
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colistin resistance superimposed to endemic carbapenem resistant klebsiella pneumoniae a rapidly evolving problem in italy november 2013 to april 2014
Eurosurveillance, 2014Co-Authors: M Monaco, M Raffone, Fabio Arena, Aurora Garciafernandez, Simona Pollini, Hajo Grundmann, Annalisa Pantosti, Tommaso Giani, Gian Maria RossoliniAbstract:Consecutive non-replicate clinical isolates (n=191) of carbapenem non-susceptible Enterobacteriaceae were collected from 21 hospital laboratories across Italy from November 2013 to April 2014 as part of the European Survey on Carbapenemase-producing Enterobacteriaceae (EuSCAPE) project. Klebsiella pneumoniae Carbapenemase-producing K. pneumoniae (KPC-KP) represented 178 (93%) isolates with 76 (43%) respectively resistant to colistin, a key drug for treating carbapenamase-producing Enterobacteriaceae. KPC-KP colistin-resistant isolates were detected in all participating laboratories. This underscores a concerning evolution of colistin resistance in a setting of high KPC-KP endemicity.
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colistin resistance superimposed to endemic carbapenem resistant klebsiella pneumoniae a rapidly evolving problem in italy november 2013 to april 2014
Eurosurveillance, 2014Co-Authors: M Monaco, M Raffone, Fabio Arena, Aurora Garciafernandez, Simona Pollini, Hajo Grundmann, Annalisa Pantosti, Tommaso Giani, Gian Maria RossoliniAbstract:Consecutive non-replicate clinical isolates (n=191) of carbapenem non-susceptible Enterobacteriaceae were collected from 21 hospital laboratories across Italy from November 2013 to April 2014 as part of the European Survey on Carbapenemase-producing Enterobacteriaceae (EuSCAPE) project. Klebsiella pneumoniae Carbapenemase-producing K. pneumoniae (KPC-KP) represented 178 (93%) isolates with 76 (43%) respectively resistant to colistin, a key drug for treating carbapenamase-producing Enterobacteriaceae. KPC-KP colistin-resistant isolates were detected in all participating laboratories. This underscores a concerning evolution of colistin resistance in a setting of high KPC-KP endemicity.
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Rapid evolution and spread of Carbapenemases among Enterobacteriaceae in Europe
Clinical Microbiology and Infection, 2012Co-Authors: Rafael Cantón, Marek Gniadkowski, Yaniv Carmeli, C. G. Giske, Thierry Naas, Murat Akova, Vivi Miriagou, Youri Glupczynski, David M. Livermore, Gian Maria RossoliniAbstract:Plasmid-acquired Carbapenemases in Enterobacteriaceae, which were first discovered in Europe in the 1990s, are now increasingly being identified at an alarming rate. Although their hydrolysis spectrum may vary, they hydrolyse most β-lactams, including carbapenems. They are mostly of the KPC, VIM, NDM and OXA-48 types. Their prevalence in Europe as reported in 2011 varies significantly from high (Greece and Italy) to low (Nordic countries). The types of Carbapenemase vary among countries, partially depending on the cultural/population exchange relationship between the European countries and the possible reservoirs of each Carbapenemase. Carbapenemase producers are mainly identified among Klebsiella pneumoniae and Escherichia coli, and still mostly in hospital settings and rarely in the community. Although important nosocomial outbreaks with Carbapenemase-producing Enterobacteriaceae have been extensively reported, many new cases are still related to importation from a foreign country. Rapid identification of colonized or infected patients and screening of carriers is possible, and will probably be effective for prevention of a scenario of endemicity, as now reported for extended-spectrum β-lactamase (mainly CTX-M) producers in all European countries.
Laurent Dortet - One of the best experts on this subject based on the ideXlab platform.
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a multiplex lateral flow immunoassay for the rapid identification of ndm kpc imp and vim type and oxa 48 like Carbapenemase producing enterobacteriaceae
Journal of Antimicrobial Chemotherapy, 2018Co-Authors: Herve Boutal, Laurent Dortet, Sandrine Bernabeu, Anais Vogel, Karine Devilliers, Elodie Creton, Garence Cotellon, Marc Plaisance, Saoussen Oueslati, Agnes B JoussetAbstract:Objectives: The global spread of Carbapenemase-producing Enterobacteriaceae represents a substantial challenge in clinical practice and rapid and reliable detection of these organisms is essential. The aim of this study was to develop and validate a lateral flow immunoassay (Carba5) for the detection of the five main Carbapenemases (KPC-, NDM-, VIM- and IMP-type and OXA-48-like). Methods: Carba5 was retrospectively and prospectively evaluated using 296 enterobacterial isolates from agar culture. An isolated colony was suspended in extraction buffer and then loaded on the manufactured Carba5. Results: All 185 isolates expressing a Carbapenemase related to one of the Carba5 targets were correctly and unambiguously detected in <15 min. All other isolates gave negative results except those producing OXA-163 and OXA-405, which are considered low-activity Carbapenemases. No cross-reaction was observed with non-targeted Carbapenemases, ESBLs, AmpCs or oxacillinases (OXA-1, -2, -9 and -10). Overall, this assay reached 100% sensitivity and 95.3% (retrospectively) to 100% (prospectively) specificity. Conclusions: Carba5 is efficient, rapid and easy to implement in the routine workflow of a clinical microbiology laboratory for confirmation of the five main Carbapenemases encountered in Enterobacteriaceae.
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a multiplex lateral flow immunoassay for the rapid identification of ndm kpc imp and vim type and oxa 48 like Carbapenemase producing enterobacteriaceae
Journal of Antimicrobial Chemotherapy, 2018Co-Authors: Herve Boutal, Laurent Dortet, Sandrine Bernabeu, Anais Vogel, Karine Devilliers, Elodie Creton, Garence Cotellon, Marc Plaisance, Saoussen Oueslati, Agnes B JoussetAbstract:Objectives The global spread of Carbapenemase-producing Enterobacteriaceae represents a substantial challenge in clinical practice and rapid and reliable detection of these organisms is essential. The aim of this study was to develop and validate a lateral flow immunoassay (Carba5) for the detection of the five main Carbapenemases (KPC-, NDM-, VIM- and IMP-type and OXA-48-like).
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A multiplex lateral flow immunoassay for the rapid identification of NDM-, KPC-, IMP- and VIM-type and OXA-48-like Carbapenemase-producing enterobacteriaceae
Journal of Antimicrobial Chemotherapy, 2018Co-Authors: Herve Boutal, Laurent Dortet, Sandrine Bernabeu, Anais Vogel, Karine Devilliers, Elodie Creton, Garence Cotellon, Marc Plaisance, Saoussen Oueslati, Agnes B JoussetAbstract:Objectives: The global spread of Carbapenemase-producing Enterobacteriaceae represents a substantial challenge in clinical practice and rapid and reliable detection of these organisms is essential. The aim of this study was to develop and validate a lateral flow immunoassay (Carba5) for the detection of the five main Carbapenemases (KPC-, NDM-, VIM-and IMP-type and OXA-48-like). Methods: Carba5 was retrospectively and prospectively evaluated using 296 enterobacterial isolates from agar culture. An isolated colony was suspended in extraction buffer and then loaded on the manufactured Carba5. Results: All 185 isolates expressing a Carbapenemase related to one of the Carba5 targets were correctly and unambiguously detected in
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evaluation of the β carba test a colorimetric test for the rapid detection of Carbapenemase activity in gram negative bacilli
Journal of Antimicrobial Chemotherapy, 2017Co-Authors: Sandrine Bernabeu, Laurent Dortet, Thierry NaasAbstract:Objectives: There is an urgent need for accurate and fast diagnostic tests to identify Carbapenemase-producing bacteria. Here, we have evaluated a novel colorimetric test (the β-CARBA™ test; Bio-Rad) to detect Carbapenemase-producing Gram-negative bacilli from cultured colonies. Methods: The performance of the β-CARBA™ test was compared with that of the Carba NP test (or the CarbAcineto NP test) and RAPIDEC ® CARBA NP (bioMerieux) using a collection of 290 isolates with characterized β-lactamase content. This collection included 199 Carbapenemase producers (121 Enterobacteriaceae, 36 Pseudomonas and 42 Acinetobacter baumannii ) and 91 non-Carbapenemase producers (55 Enterobacteriaceae, 20 Pseudomonas and 16 A. baumannii ). Results: The β-CARBA™ test correctly detected 84.9% of the Carbapenemase producers, including all KPC and IMP, 96.4% of VIM, 85.3% of NDM, 80.5% of OXA-48-like and 91.2% of A. baumannii -related OXA Carbapenemases (OXA-23, OXA-40, OXA-58, OXA-143 and overexpressed OXA-51). All rare metallo-β-lactamases (SPM, AIM, GIM, DIM and SIM) were detected. Importantly, all non-KPC Ambler class A Carbapenemases were not detected, including GES variants with Carbapenemase activity ( n = 6), IMI ( n = 3), NMC-A ( n = 1), SME ( n = 2), FRI-1 ( n = 1) and BIC-1 ( n = 1). All non-Carbapenemase producers gave a negative result except with OXA-163-, OXA-405- and one TEM-3-producing Citrobacter freundii . The overall sensitivity and specificity of the β-CARBA™ test were 84.9% and 95.6%, respectively. This test is easy to perform and to interpret by non-specialized staff members. Conclusions: Despite lack of specificity towards non-KPC Ambler class A and OXA-48-like Carbapenemases, the β-CARBA™ test could complete the existing panel of tests available for the confirmation of Carbapenemases in Gram-negatives.
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Development and validation of a lateral flow immunoassay for rapid detection of NDM-producing Enterobacteriaceae
Journal of Clinical Microbiology, 2017Co-Authors: Herve Boutal, Thierry Naas, Laurent Dortet, Sandrine Bernabeu, Karine Devilliers, Saoussen Oueslati, Stéphanie Simon, Hervé VollandAbstract:The global spread of Carbapenemase-producing Enterobacteriaceae (CPE) that are often resistant to most, if not all, classes of antibiotics is a major public health concern. The NDM-1 Carbapenemase is among the most worrisome Carbapenemases given its rapid worldwide spread. We have developed and evaluated a lateral flow immunoassay (LFIA) (called the NDM LFIA) for the rapid and reliable detection of NDM-like Carbapenemase-producing Enterobacteriaceae from culture colonies. We evaluated the NDM LFIA using 175 reference enterobacterial isolates with characterized β-lactamase gene content and 74 nonduplicate consecutive carbapenem-resistant clinical isolates referred for expertise to the French National Reference Center (NRC) for Antibiotic Resistance during a 1-week period (in June 2016). The reference collection included 55 non-Carbapenemase producers and 120 Carbapenemase producers, including 27 NDM producers. All 27 NDM-like Carbapenemase producers of the reference collection were correctly detected in less than 15 min by the NDM LFIA, including 22 strains producing NDM-1, 2 producing NDM-4, 1 producing NDM-5, 1 producing NDM-7, and 1 producing NDM-9. All non-NDM-1 producers gave a negative result with the NDM LFIA. No cross-reaction was observed with Carbapenemases (VIM, IMP, NDM, KPC, and OXA-48-like), extended-spectrum β-lactamases (ESBLs) (TEM, SHV, and CTX-M), AmpCs (CMY-2, DHA-2, and ACC-1), and oxacillinases (OXA-1, -2, -9, and -10). Similarly, among the 74 referred nonduplicate consecutive clinical isolates, all 7 NDM-like producers were identified. Overall, the sensitivity and specificity of the assay were 100% for NDM-like Carbapenemase detection with strains cultured on agar. The NDM LFIA was efficient, rapid, and easy to implement in the routine workflow of a clinical microbiology laboratory for the confirmation of NDM-like Carbapenemase-producing Enterobacteriaceae.
Neil Woodford - One of the best experts on this subject based on the ideXlab platform.
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Evaluation of temocillin for phenotypic Carbapenemase screening of Escherichia coli and Salmonella enterica isolates in relation to the presence of genes encoding ESBLs and Carbapenemase production
Journal of Antimicrobial Chemotherapy, 2019Co-Authors: Lina Cavaco, Neil Woodford, Robert Hill, Frank Hansen, Shazad Mushtaq, Simon Le Hello, Rene Hendriksen, Anette Hammerum, Henrik HasmanAbstract:The expression of enzymes of the OXA-48 Carbapenemase group is difficult to detect by phenotypic methods owing to frequent low levels of carbapenem resistance and negative results with some screening methods. Temocillin has been shown to be a good option for phenotypic screening as it is hydrolysed by the OXA-48-group enzymes, whereas ESBLs, AmpC and some other Carbapenemases have a lower hydrolytic effect on this antimicrobial. However, no epidemiological cut-off for temocillin is available. OBJECTIVES: To evaluate temocillin MICs in relation to the presence or absence of genes encoding ESBLs and Carbapenemases in Escherichia coli and Salmonella enterica. METHODS: In this study, 111 E. coli and 102 S. enterica isolates, including WT and well-characterized ESBL-, AmpC- or Carbapenemase-producing isolates, were tested by three independent laboratories. MICs were determined according to the CLSI guidelines by agar dilution with the test range from 0.5 to 512 mg/L temocillin and WGS was performed and analysed with ResFinder. RESULTS: Some overlap was detected between temocillin MICs for WT and ESBL- or AmpC-producing isolates. However, isolates carrying genes encoding Carbapenemases showed a broader range of MICs for both E. coli and S. enterica. Higher MICs were observed for the OXA-48 group, VIM and some NDM-producing isolates, whereas isolates harbouring KPC enzymes showed low MICs. CONCLUSIONS: The results indicate that temocillin MICs enable phenotypic distinction between strains producing OXA-48-group enzymes and both WT susceptible and ESBL/AmpC-carrying isolates, whereas the distinction from other Carbapenemases likely requires genotypic testing.
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Carbapenemase producing enterobacteriaceae and non enterobacteriaceae from animals and the environment an emerging public health risk of our own making
Journal of Antimicrobial Chemotherapy, 2014Co-Authors: Neil Woodford, David W Wareham, Beatriz Guerra, Christopher TealeAbstract:Acquired Carbapenemases pose one of the most pressing public health threats relating to antibiotic resistance. In most countries, the number of Carbapenemase-producing bacteria from human clinical specimens is rising, and the epidemiological status of these multiresistant bacteria is progressively worsening. Furthermore, there is a growing number of reports of Carbapenemases found either in bacteria isolated from non-human sources or in Salmonella enterica subsp. enterica, a zoonotic species. However, Carbapenemases are not yet systematically sought in bacteria from non-human sources, reports of them are largely observational, and there is limited investigation of Carbapenemase-positive bacteria in animals and possible links with people who may have acted as potential sources. Active surveillance and monitoring for carbapenem-resistant bacteria in the food chain and other non-human sources is urgently needed, with an enhanced and rigorous follow-up of all positive results. The carbapenems are currently our last good defence against multiresistant Gram-negative bacteria. Our ability to limit the rise and spread of Carbapenemase producers, which occur only at basal levels in many countries at present, should serve as a key performance indicator for the success or failure of the efforts that have been called for by international organizations and governments to reduce the impact of antibiotic resistance.
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in vitro activity of temocillin against multidrug resistant clinical isolates of escherichia coli klebsiella spp and enterobacter spp and evaluation of high level temocillin resistance as a diagnostic marker for oxa 48 Carbapenemase
Journal of Antimicrobial Chemotherapy, 2014Co-Authors: Neil Woodford, Danièle Meunier, Rachel Pike, Robert Hill, Katie L HopkinsAbstract:Sir, Temocillin is a narrow-spectrum penicillin active primarily against Enterobacteriaceae and resistant to hydrolysis by penicillinases, extended-spectrum b-lactamases (ESBLs), and AmpC enzymes. It has been shown also to retain some activity in vitro against Enterobacteriaceae with KPC-type Carbapenemases, although not against bacteria with OXA-48-like non-metalloenzymes or the metalloenzymes (including IMP, NDM and VIM types). The emergence and spread of bacteria with acquired Carbapenemases raises public health concerns, and there is a clear need for reliable diagnostic tests in routine bacteriology laboratories. Simple phenotypic tests with b-lactam/b-lactamase inhibitor combinations can aid the identification of isolates with KPC-type or metallo-Carbapenemases, but cannot detect OXA-48 Carbapenemases, for which there is currently no good inhibitor. Molecular tests could overcome this shortcoming but are not an option in many laboratories. The Carba NP acido/colorimetric phenotypic test detects carbapenem hydrolysis per se, but its sensitivity is reportedly lower for OXA-48 producers than for isolates with other Carbapenemases. High-level temocillin resistance is a characteristic of many OXA-48 producers and has been included in algorithms to aid their recognition. – 8 We investigated the in vitro activity of temocillin against 2280 clinical isolates of Escherichia coli, Klebsiella spp. and Enterobacter spp. submitted to Public Health England’s Antimicrobial Resistance and Healthcare Associated Infections (AMRHAI) Reference Unit for the investigation of unusual resistance between January 2012 and April 2013. They included 1029 producers of KPC, OXA-48 or IMP, NDM and VIM Carbapenemases. We also evaluated the diagnostic potential of high-level temocillin resistance (MIC ≥128 mg/L) to predict the production of an OXA-48 Carbapenemase. Temocillin (Eumedica, Brussels, Belgium) was tested in the range 1–128 mg/L using agar dilution methodology; MICs were interpreted using BSAC breakpoints (susceptibility: systemic infections, MICs≤8 mg/L; urinary infections, MICs≤32 mg/L). Isolates were also tested against AMRHAI’s standard Gram-negative antibiotic panel, which includes ertapenem, imipenem (tested+EDTA to indicate likely metallo-Carbapenemase producers) and meropenem. Carbapenemase production was sought using in-house PCRs and/or a commercial microarray (Check-MDR CT102; Check-Points, Wageningen, The Netherlands). The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of temocillin MIC ≥128 mg/L as an indicator of OXA-48 Carbapenemase production were determined using an online tool (http://www. medcalc.org/calc/diagnostic_test.php). Temocillin MIC distributions for the 2280 isolates are shown in Table 1. These were submissions to a reference laboratory so were biased towards multiresistance, with proportions of ‘unusual’ isolates far higher than would be observed in most diagnostic laboratories. Allowing for the fact that a larger proportion of Klebsiella spp. isolates produced Carbapenemases (760/1133; 67.1%) than did isolates of E. coli (206/701; 29.4%) or Enterobacter spp. (63/446; 14.1%), no major differences were observed in the MIC distributions of temocillin for the three genera. Overall, 920 (40.4%) isolates were susceptible to temocillin at the systemic breakpoint (MICs ≤8 mg/L); those considered resistant included 329/669 (49.2%) KPC producers, 671/1251 (53.6%) isolates without Carbapenemases, and all 360 (100%) isolates with other types of Carbapenemase (OXA-48, IMP, NDM or VIM). At the higher, urinary breakpoint, 1758 (77.1%) isolates were susceptible to temocillin (MICs ≤32 mg/L), including 628 (93.4%) KPC producers and 1098 (87.8%) isolates without Carbapenemases. However, 328 (91.4%) isolates with other Carbapenemases (OXA-48, IMP, NDM or VIM) remained resistant with MICs .32 mg/L. Temocillin MICs ≥128 mg/L were observed for 355 (15.6%) isolates overall, most (292; 82.3%) of which had a Carbapenemase. However, 63 isolates (17.7%; 30 E. coli, 22 Klebsiella spp. and 11 Enterobacter spp.) lacked these mechanisms and the nature of their high-level temocillin resistance warrants further investigation. Many of them had complex b-lactam antibiograms—for example, 57 were also non-susceptible to ertapenem (MICs .0.5 mg/L)—consistent with ESBL/AmpC production combined with impermeability (data not shown). High-level temocillin resistance (MICs ≥128 mg/L) was recorded for 98/108 (90.7%) OXA-48 producers compared with 257/2172 (11.8%) isolates lacking OXA-48 or with 72/1920 (3.8%) isolates lacking a metallo-Carbapenemase. Based on these proportions, the detection of high-level resistance was not a sufficiently robust marker when used as the sole criterion to predict the presence of an OXA-48-like Carbapenemase; the PPV of the test was only 27.6% (Table 2). However, if combined with an absence of imipenem/EDTA synergy (i.e. when there was no phenotypic evidence of metallo-Carbapenemase production), the PPV of high-level temocillin resistance predicting OXA-48
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comparison of bd phoenix vitek 2 and microscan automated systems for detection and inference of mechanisms responsible for carbapenem resistance in enterobacteriaceae
Journal of Clinical Microbiology, 2010Co-Authors: Neil Woodford, Anne Eastaway, Michael Ford, Alistair Leanord, Chloe Keane, Reinhard M Quayle, Jane A Steer, Jiancheng Zhang, David M. LivermoreAbstract:We assessed the ability of three commercial systems to infer carbapenem resistance mechanisms in 39 Carbapenemase-producing and 16 other carbapenem-resistant Enterobacteriaceae. The sensitivity/specificity values for “flagging” a likely Carbapenemase were 100%/0% (BD Phoenix), 82 to 85%/6 to 19% (MicroScan), and 74%/38% (Vitek 2), respectively. OXA-48 producers were poorly detected, but all systems reliably detected isolates with KPC and most with metallo-Carbapenemases.
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Carbapenemases: a problem in waiting?
Current opinion in microbiology, 2000Co-Authors: David M. Livermore, Neil WoodfordAbstract:Carbapenems are stable to most prevalent β-lactamases, and chromosomal Carbapenemases are restricted to Stenotrophomonas maltophilia, to a few Bacteroides fragilis, and to rare pathogens. Nevertheless, an acquired metallo-β-lactamase called IMP-1 is beginning to emerge in Pseudomonas aeruginosa and Enterobacteriaceae isolates in Japan, and has also been found in isolates from Singapore. Furthermore, IMP-producing Acinetobacter spp. have been identified in Italy and Hong Kong. Recently a second group of acquired metallo-Carbapenemases, the VIM types, has been recorded from P. aeruginosa isolates in five Eurasian countries. Weak Carbapenemases belonging to molecular class D are emerging in A. baumannii world-wide, with two sub-groups apparent. A few acquired Carbapenemases belonging to molecular class A also have been reported. Finally it has also been shown that enzymes with feeble Carbapenemase activity (e.g. AmpC types and some SHV enzymes) may confer resistance in exceptionally impermeable strains; counterwise, even potent Carbapenemases, such as IMP-1, may only give a small reduction in susceptibility in Enterobacteriaceae that lack permeability lesions. Is the emergence of Carbapenemase a problem waiting to happen?