Skip to main content
Top
Published in: BMC Infectious Diseases 1/2017

Open Access 01-12-2017 | Research article

Development of an algorithm for phenotypic screening of carbapenemase-producing Enterobacteriaceae in the routine laboratory

Authors: Jérôme Robert, Alix Pantel, Audrey Merens, Elodie Meiller, Jean-Philippe Lavigne, Marie-Hélène Nicolas-Chanoine, on behalf of ONERBA’s carbapenem resistance study group

Published in: BMC Infectious Diseases | Issue 1/2017

Login to get access

Abstract

Background

Carbapenemase-producing Enterobacteriaceae (CPE) are difficult to identify among carbapenem non-susceptible Enterobacteriaceae (NSE). We designed phenotypic strategies giving priority to high sensitivity for screening putative CPE before further testing.

Methods

Presence of carbapenemase-encoding genes in ertapenem NSE (MIC > 0.5 mg/l) consecutively isolated in 80 French laboratories between November 2011 and April 2012 was determined by the Check-MDR-CT103 array method. Using the Mueller-Hinton (MH) disk diffusion method, clinical diameter breakpoints of carbapenems other than ertapenem, piperazicillin+tazobactam, ticarcillin+clavulanate and cefepime as well as diameter cut-offs for these antibiotics and temocillin were evaluated alone or combined to determine their performances (sensitivity, specificity, positive and negative likelihood ratios) for identifying putative CPE among these ertapenem-NSE isolates. To increase the screening specificity, these antibiotics were also tested on cloxacillin-containing MH when carbapenem NSE isolates belonged to species producing chromosomal cephalosporinase (AmpC) but Escherichia coli.

Results

Out of the 349 ertapenem NSE, 52 (14.9%) were CPE, including 39 producing OXA-48 group carbapenemase, eight KPC and five MBL. A screening strategy based on the following diameter cut offs, ticarcillin+clavulanate <15 mm, temocillin <15 mm, meropenem or imipenem <22 mm, and cefepime <26 mm, showed 100% sensitivity and 68.1% specificity with the better likelihood ratios combination. The specificity increased when a diameter cut-off <32 mm for imipenem (76.1%) or meropenem (78.8%) further tested on cloxacillin-containing MH was added to the previous strategy for AmpC-producing isolates.

Conclusion

The proposed strategies that allowed for increasing the likelihood of CPE among ertapenem-NSE isolates should be considered as a surrogate for carbapenemase production before further CPE confirmatory testing.
Appendix
Available only for authorised users
Literature
1.
go back to reference Carmeli Y, Akova M, Cornaglia G, Daikos GL, Garau J, Harbarth S, et al. Controlling the spread of carbapenemase-producing Gram-negatives: therapeutic approach and infection control. Clin Microbiol Infect. 2010;16:102–11.CrossRefPubMed Carmeli Y, Akova M, Cornaglia G, Daikos GL, Garau J, Harbarth S, et al. Controlling the spread of carbapenemase-producing Gram-negatives: therapeutic approach and infection control. Clin Microbiol Infect. 2010;16:102–11.CrossRefPubMed
2.
go back to reference Fournier S, Monteil C, Lepainteur M, Richard C, Brun-Buisson C, Jarlier V, Ap-Hp Outbreaks Control Group C. Long-term control of carbapenemase-producing Enterobacteriaceae at the scale of a large French multihospital institution: a nine-year experience, France, 2004 to 2012. Euro Surveill. 2014;19:20802.CrossRefPubMed Fournier S, Monteil C, Lepainteur M, Richard C, Brun-Buisson C, Jarlier V, Ap-Hp Outbreaks Control Group C. Long-term control of carbapenemase-producing Enterobacteriaceae at the scale of a large French multihospital institution: a nine-year experience, France, 2004 to 2012. Euro Surveill. 2014;19:20802.CrossRefPubMed
3.
go back to reference Huang T-D, Berhin C, Bogaerts P, Glupczynski Y. Prevalence and mechanisms of resistance to carbapenems in Enterobacteriaceae isolates from 24 hospitals in Belgium. J Antimicrob Chemother. 2013;68:1832–7.CrossRefPubMed Huang T-D, Berhin C, Bogaerts P, Glupczynski Y. Prevalence and mechanisms of resistance to carbapenems in Enterobacteriaceae isolates from 24 hospitals in Belgium. J Antimicrob Chemother. 2013;68:1832–7.CrossRefPubMed
4.
go back to reference Lee EH, Nicolas MH, Kitzis MD, Pialoux G, Collatz E, Gutmann L. Association of two resistance mechanisms in a clinical isolate of Enterobacter cloacae with high-level resistance to imipenem. Antimicrob Agents Chemother. 1991;35:1093–8.CrossRefPubMedPubMedCentral Lee EH, Nicolas MH, Kitzis MD, Pialoux G, Collatz E, Gutmann L. Association of two resistance mechanisms in a clinical isolate of Enterobacter cloacae with high-level resistance to imipenem. Antimicrob Agents Chemother. 1991;35:1093–8.CrossRefPubMedPubMedCentral
5.
go back to reference Robert J, Pantel A, Mérens A, Lavigne J-P, Nicolas-Chanoine M-H. Incidence rates of carbapenemase-producing Enterobacteriaceae clinical isolates in France: a prospective nationwide study in 2011–12. J Antimicrob Chemother. 2014;69:2706–12.CrossRefPubMed Robert J, Pantel A, Mérens A, Lavigne J-P, Nicolas-Chanoine M-H. Incidence rates of carbapenemase-producing Enterobacteriaceae clinical isolates in France: a prospective nationwide study in 2011–12. J Antimicrob Chemother. 2014;69:2706–12.CrossRefPubMed
6.
go back to reference Tenover FC, Canton R, Kop J, Chan R, Ryan J, Weir F, et al. Detection of colonization by carbapenemase-producing Gram-negative Bacilli in patients by use of the Xpert MDRO assay. J Clin Microbiol. 2013;51:3780–7.CrossRefPubMedPubMedCentral Tenover FC, Canton R, Kop J, Chan R, Ryan J, Weir F, et al. Detection of colonization by carbapenemase-producing Gram-negative Bacilli in patients by use of the Xpert MDRO assay. J Clin Microbiol. 2013;51:3780–7.CrossRefPubMedPubMedCentral
7.
go back to reference Knox J, Jadhav S, Sevior D, Agyekum A, Whipp M, Waring L, et al. Phenotypic detection of carbapenemase-producing Enterobacteriaceae using MALDI-TOF MS and the Carba NP test. J Clin Microbiol. 2014;52:4075–7.CrossRefPubMedPubMedCentral Knox J, Jadhav S, Sevior D, Agyekum A, Whipp M, Waring L, et al. Phenotypic detection of carbapenemase-producing Enterobacteriaceae using MALDI-TOF MS and the Carba NP test. J Clin Microbiol. 2014;52:4075–7.CrossRefPubMedPubMedCentral
8.
9.
go back to reference Osei Sekyere J, Govinden U, Essack SY. Review of established and innovative detection methods for carbapenemase-producing Gram-negative bacteria. J Appl Microbiol. 2015;119:1219–33.CrossRefPubMed Osei Sekyere J, Govinden U, Essack SY. Review of established and innovative detection methods for carbapenemase-producing Gram-negative bacteria. J Appl Microbiol. 2015;119:1219–33.CrossRefPubMed
10.
go back to reference Cohen Stuart J, Leverstein-Van Hall MA. Guideline for phenotypic screening and confirmation of carbapenemases in Enterobacteriaceae. Int J Antimicrob Agents. 2010;36:205–10.CrossRefPubMed Cohen Stuart J, Leverstein-Van Hall MA. Guideline for phenotypic screening and confirmation of carbapenemases in Enterobacteriaceae. Int J Antimicrob Agents. 2010;36:205–10.CrossRefPubMed
11.
go back to reference Dortet L, Bréchard L, Cuzon G, Poirel L, Nordmann P. Strategy for a rapid detection of carbapenemase-producing Enterobacteriaceae. Antimicrob Agents Chemother. 2014;58:2441–5.CrossRefPubMedPubMedCentral Dortet L, Bréchard L, Cuzon G, Poirel L, Nordmann P. Strategy for a rapid detection of carbapenemase-producing Enterobacteriaceae. Antimicrob Agents Chemother. 2014;58:2441–5.CrossRefPubMedPubMedCentral
12.
go back to reference Doyle D, Peirano G, Lascols C, Lloyd T, Church DL, Pitout JDD. Laboratory detection of Enterobacteriaceae that produce carbapenemases. J Clin Microbiol. 2012;50:3877–80.CrossRefPubMedPubMedCentral Doyle D, Peirano G, Lascols C, Lloyd T, Church DL, Pitout JDD. Laboratory detection of Enterobacteriaceae that produce carbapenemases. J Clin Microbiol. 2012;50:3877–80.CrossRefPubMedPubMedCentral
13.
go back to reference Hrabák J, Chudáčková E, Papagiannitsis CC. Detection of carbapenemases in Enterobacteriaceae: a challenge for diagnostic microbiological laboratories. Clin Microbiol Infect. 2014;20:839–53.CrossRefPubMed Hrabák J, Chudáčková E, Papagiannitsis CC. Detection of carbapenemases in Enterobacteriaceae: a challenge for diagnostic microbiological laboratories. Clin Microbiol Infect. 2014;20:839–53.CrossRefPubMed
14.
go back to reference Huang T-D, Poirel L, Bogaerts P, Berhin C, Nordmann P, Glupczynski Y. Temocillin and piperacillin/tazobactam resistance by disc diffusion as antimicrobial surrogate markers for the detection of carbapenemase-producing Enterobacteriaceae in geographical areas with a high prevalence of OXA-48 producers. J Antimicrob Chemother. 2014;69:445–50.CrossRefPubMed Huang T-D, Poirel L, Bogaerts P, Berhin C, Nordmann P, Glupczynski Y. Temocillin and piperacillin/tazobactam resistance by disc diffusion as antimicrobial surrogate markers for the detection of carbapenemase-producing Enterobacteriaceae in geographical areas with a high prevalence of OXA-48 producers. J Antimicrob Chemother. 2014;69:445–50.CrossRefPubMed
15.
go back to reference van Dijk K, Voets GM, Scharringa J, Voskuil S, Fluit AC, Rottier WC, et al. A disc diffusion assay for detection of class A, B and OXA-48 carbapenemases in Enterobacteriaceae using phenyl boronic acid, dipicolinic acid and temocillin. Clin Microbiol Infect. 2014;20:345–9.CrossRefPubMed van Dijk K, Voets GM, Scharringa J, Voskuil S, Fluit AC, Rottier WC, et al. A disc diffusion assay for detection of class A, B and OXA-48 carbapenemases in Enterobacteriaceae using phenyl boronic acid, dipicolinic acid and temocillin. Clin Microbiol Infect. 2014;20:345–9.CrossRefPubMed
16.
go back to reference European Committee for Antimicrobial Susceptibility Testing (EUCAST) of the European Society of Clinical Microbiology and Infectious Diseases (ESCMID). Determination of minimum inhibitory concentrations (MICs) of antibacterial agents by broth dilution. Clin Microbiol Infect. 2003;9:ix–xv. European Committee for Antimicrobial Susceptibility Testing (EUCAST) of the European Society of Clinical Microbiology and Infectious Diseases (ESCMID). Determination of minimum inhibitory concentrations (MICs) of antibacterial agents by broth dilution. Clin Microbiol Infect. 2003;9:ix–xv.
18.
go back to reference Woodford N, Pike R, Meunier D, Loy R, Hill R, Hopkins KL. 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. J Antimicrob Chemother. 2014;69:564–7.CrossRefPubMed Woodford N, Pike R, Meunier D, Loy R, Hill R, Hopkins KL. 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. J Antimicrob Chemother. 2014;69:564–7.CrossRefPubMed
19.
go back to reference Caraguel CGB, Vanderstichel R. The two-step Fagan’s nomogram: ad hoc interpretation of a diagnostic test result without calculation. Evid Based Med. 2013;18:125–8.CrossRefPubMed Caraguel CGB, Vanderstichel R. The two-step Fagan’s nomogram: ad hoc interpretation of a diagnostic test result without calculation. Evid Based Med. 2013;18:125–8.CrossRefPubMed
20.
go back to reference Garrec H, Drieux-Rouzet L, Golmard J-L, Jarlier V, Robert J. Comparison of nine phenotypic methods for detection of Extended-spectrum β-lactamase production by Enterobacteriaceae. J Clin Microbiol. 2011;49:1048–57.CrossRefPubMedPubMedCentral Garrec H, Drieux-Rouzet L, Golmard J-L, Jarlier V, Robert J. Comparison of nine phenotypic methods for detection of Extended-spectrum β-lactamase production by Enterobacteriaceae. J Clin Microbiol. 2011;49:1048–57.CrossRefPubMedPubMedCentral
21.
go back to reference Biggerstaff BJ. Comparing diagnostic tests: a simple graphic using likelihood ratios. Stat Med. 2000;19:649–63.CrossRefPubMed Biggerstaff BJ. Comparing diagnostic tests: a simple graphic using likelihood ratios. Stat Med. 2000;19:649–63.CrossRefPubMed
22.
go back to reference Dortet L, Cuzon G, Plésiat P, Naas T. Prospective evaluation of an algorithm for the phenotypic screening of carbapenemase-producing Enterobacteriaceae. J Antimicrob Chemother. 2016;71:135–40.CrossRefPubMed Dortet L, Cuzon G, Plésiat P, Naas T. Prospective evaluation of an algorithm for the phenotypic screening of carbapenemase-producing Enterobacteriaceae. J Antimicrob Chemother. 2016;71:135–40.CrossRefPubMed
23.
go back to reference Dortet L, Cuzon G, Nordmann P. Dissemination of carbapenemase-producing Enterobacteriaceae in France, 2012. J Antimicrob Chemother. 2014;69:623–7.CrossRefPubMed Dortet L, Cuzon G, Nordmann P. Dissemination of carbapenemase-producing Enterobacteriaceae in France, 2012. J Antimicrob Chemother. 2014;69:623–7.CrossRefPubMed
24.
go back to reference Picão RC, Santos AF, Nicoletti AG, Furtado GH, Gales AC. Detection of GES-5-producing Klebsiella pneumoniae in Brazil. J Antimicrob Chemother. 2010;65:796–7.CrossRefPubMed Picão RC, Santos AF, Nicoletti AG, Furtado GH, Gales AC. Detection of GES-5-producing Klebsiella pneumoniae in Brazil. J Antimicrob Chemother. 2010;65:796–7.CrossRefPubMed
26.
go back to reference Pasteran F, Veliz O, Ceriana P, Lucero C, Rapoport M, Albornoz E, Gomez S, Corso A. Evaluation of the Blue-Carba test for rapid detection of carbapenemases in gram-negative bacilli. J Clin Microbiol. 2015;53:1996–8.CrossRefPubMedPubMedCentral Pasteran F, Veliz O, Ceriana P, Lucero C, Rapoport M, Albornoz E, Gomez S, Corso A. Evaluation of the Blue-Carba test for rapid detection of carbapenemases in gram-negative bacilli. J Clin Microbiol. 2015;53:1996–8.CrossRefPubMedPubMedCentral
Metadata
Title
Development of an algorithm for phenotypic screening of carbapenemase-producing Enterobacteriaceae in the routine laboratory
Authors
Jérôme Robert
Alix Pantel
Audrey Merens
Elodie Meiller
Jean-Philippe Lavigne
Marie-Hélène Nicolas-Chanoine
on behalf of ONERBA’s carbapenem resistance study group
Publication date
01-12-2017
Publisher
BioMed Central
Published in
BMC Infectious Diseases / Issue 1/2017
Electronic ISSN: 1471-2334
DOI
https://doi.org/10.1186/s12879-016-2174-y

Other articles of this Issue 1/2017

BMC Infectious Diseases 1/2017 Go to the issue
Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.