Skip to main content
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems
  • Log in
  • My alerts
  • My Cart

Main menu

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • COVID-19 Special Collection
    • Archive
    • Minireviews
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About JCM
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems

User menu

  • Log in
  • My alerts
  • My Cart

Search

  • Advanced search
Journal of Clinical Microbiology
publisher-logosite-logo

Advanced Search

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • COVID-19 Special Collection
    • Archive
    • Minireviews
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About JCM
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
Letter to the Editor

Rapid Detection of Carbapenemases in Enterobacteriaceae: Evaluation of the Resist-3 O.K.N. (OXA-48, KPC, NDM) Lateral Flow Multiplexed Assay

David W. Wareham, Muhd Haziq F. Abdul Momin
Karen C. Carroll, Editor
David W. Wareham
aAntimicrobial Research Group, Blizard Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom
bDivision of Infection, Barts Health NHS Trust, London, United Kingdom
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Muhd Haziq F. Abdul Momin
aAntimicrobial Research Group, Blizard Institute, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Karen C. Carroll
The Johns Hopkins University School of Medicine
Roles: Editor
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1128/JCM.02471-16
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

LETTER

The identification, treatment, and control of carbapenem-resistant Enterobacteriaceae (CRE) infections are a major challenge for health care institutions and diagnostic laboratories worldwide. Those producing plasmid-mediated OXA-48, KPC, NDM, or VIM-like carbapenemases (CPE) are most concerning, being frequently involved in nosocomial outbreaks that are difficult and very costly to manage (1). A simple and rapid test able to provide sensitive and specific identification of CRE and also the carbapenemase present is critical in any strategy aimed at addressing this problem.

Numerous phenotypic (MIC determination, disc diffusion, selective culture media, acidimetric) and genotypic (PCR amplification, microarrays, DNA sequencing) methods have been used in the laboratory isolation, diagnosis, and confirmation of CRE (2). Recently, two novel immunochromatographic lateral flow assays (Coris BioConcept, Gembloux, Belgium) were developed for the specific detection of OXA-48 (OXA-48 K-SeT) and KPC-like (KPC K-SeT) carbapenemase producers. These assays have been evaluated in multiple laboratories, using diverse sets of organisms (Escherichia coli, Klebsiella, Enterobacter, Serratia, Providencia, Pseudomonas spp.) carrying multiple β-lactamase, OXA-48 (OXA-48, -162, -181, -204, -232, -242), and KPC (KPC-2/3/4) allelic variants (3, 4, 5, 6). Both have a reported sensitivity and specificity of 100% compared to the results of molecular detection of carbapenemase genes as the gold standard (3, 4, 5, 6). They have also been shown to be compatible with organisms recovered from most unsupplemented, selective, solid, and liquid culture media currently in use in diagnostic laboratories and with bacteria taken directly from positive blood culture bottles (BacT/Alert; bioMérieux, Macy L'Etoile, France) or culture-positive urinary samples (3, 4). A modification of this system, Resist-3 O.K.N. (Coris BioConcept) designed for the simultaneous detection of OXA-48, KPC, and NDM-like enzymes using a single disposable cartridge, has now been manufactured. Here we assessed the ability of the Resist-3 O.K.N. assay to detect OXA-48, KPC, and NDM coproduction in a collection of 112 nonreplicate well-defined CRE isolates received in our laboratories.

All isolates were carbapenem-resistant Enterobacteriaceae (ertapenem MIC, >1 μg/ml) recovered from routine clinical samples submitted to Barts Health NHS Trust or the Antimicrobial Research Laboratory at Queen Mary University of London between January 2011 and December 2016. Identification was performed using the Bruker matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry system (Bruker UK Ltd., Coventry, UK), with carbapenem MICs determined using MicroScan WalkAway Negative Combo 36 panels (Siemens Healthcare Diagnostics, Deerfield, IL) and confirmed with Etest gradient strips (bioMérieux). Multiplex PCRs were used to confirm the presence of class A (KPC, BKC, SME, VEB, PER, GES), B (IMP, VIM, NDM, SIM, SPM, DIM, GIM, KHM, FIM, AIM, DIM, TMB, FRI), and D (OXA-like) carbapenemase genes, and all allelic variants identified were confirmed by Sanger sequencing of the entire coding regions (7, 8). The collection consisted of carbapenemase-producing Klebsiella pneumoniae (n = 57), E. coli (n = 17), Enterobacter spp. (n = 14), and Providencia stuartii (n = 3) and single isolates of Citrobacter koseri, Morganella morganii, and Proteus mirabilis (Table 1). Carbapenemases produced by the isolates were identified as KPC-2/4, NDM-1/5/7, VIM-1/4, IMP-1, OXA-48, and OXA-232. Nine isolates produced 2 carbapenemases, either OXA-48 in combination with NDM-1 (n = 6) or KPC-2 in combination with VIM-1 (n = 3).

View this table:
  • View inline
  • View popup
TABLE 1

Detection of carbapenemase production using the Resist-3 O.K.N. assay

Detection of OXA-48, KPC, and NDM using the Resist-3 O.K.N. cassettes was carried out according to the manufacturer's protocol. Bacteria were grown for 18 h at 37°C on Mueller-Hinton II agar plates (Oxoid), and a single colony was emulsified in 5 drops of the lysis buffer. Cassettes were loaded with 3 drops of lysate and read within 5 min. Six carbapenem-susceptible Gram-negative type strains (K. pneumoniae NCTC 9633, E. coli NCTC 12241, Enterobacter cloacae NCTC 13380, Enterobacter aerogenes NCTC 9375, Pseudomonas aeruginosa ATCC 27852, Acinetobacter baumannii ATCC 19606) were used as negative controls. Eighteen additional Enterobacteriaceae isolates (K. pneumoniae [n = 8] or Klebsiella oxytoca [n = 1], E. coli [n = 4], E. cloacae [n = 2] or E. aerogenes [n = 2], Serratia marcescens [n = 1]) with phenotypic resistance to ertapenem but without a known carbapenemase were also used as negative controls.

There was complete agreement between the carbapenemases detected by PCR and the results obtained with Resist-3 O.K.N. (Table 1). The lateral flow device was able to correctly identify and differentiate OXA, KPC, and NDM production among all CRE species and enzyme variants tested, including those carrying more than one carbapenemase. No cross-reactions were observed for strains with carbapenem resistance due to IMP-1 or VIM-1/4 (Table 1) with any of the susceptible type strains or with the 18 CRE that had no OXA, KPC, or NDM carbapenemase detectable by PCR (Fig. 1).

FIG 1
  • Open in new tab
  • Download powerpoint
FIG 1

Detection of OXA-48, KPC, and NDM-like carbapenemases in isolates producing single and dual enzymes using the Resist-3 O.K.N. assay. NEG, negative; POS, positive.

We found the Resist-3 O.K.N. assay to be 100% sensitive and specific in the detection and differentiation of OXA-48, KPC, and NDM-like carbapenemases among CRE recently referred to our laboratory. Although the assay does not currently extend to the detection of VIM or IMP-like metallo-β-lactamases, the ability to detect 3 of the 5 most prevalent and transmissible carbapenemases found worldwide is a significant advantage. The ease of use, speed, and low cost (less than $15) of the cassettes make it attractive as a diagnostic tool for use in the management of CRE outbreaks. Its role either as a primary-screening, confirmatory, or rapid point-of-care test should be assessed further, ideally prospectively in the setting of a polyclonal nosocomial CRE outbreak.

ACKNOWLEDGMENTS

Resist-3 O.K.N. assays were supplied free of charge for evaluation by BioConnections (Knypersley, UK). No other specific funding was used to undertake this study, as it was performed as part of our routine activities.

We declare no conflict of interest and have no association with Coris BioConcept.

  • Copyright © 2017 American Society for Microbiology.

All Rights Reserved .

REFERENCES

  1. 1.↵
    1. Otter JA,
    2. Burgess P,
    3. Davies F,
    4. Mookerjee S,
    5. Singleton J,
    6. Gilchrist M,
    7. Parsons D,
    8. Brannigan ET,
    9. Robotham J,
    10. Holmes AH
    . 13October2016. Counting the cost of an outbreak of carbapenemase-producing Enterobacteriaceae: an economic evaluation from a hospital perspective. Clin Microbiol Infect doi:10.1016/j.cmi.2016.10.005.
    OpenUrlCrossRef
  2. 2.↵
    1. Lutgring JD,
    2. Limbago BM
    . 2016. The problem of carbapenemase-producing-carbapenem-resistant-Enterobacteriaceae detection. J Clin Microbiol54:529–534. doi:10.1128/JCM.02771-15.
    OpenUrlAbstract/FREE Full Text
  3. 3.↵
    1. Wareham DW,
    2. Shah R,
    3. Betts JW,
    4. Phee LM,
    5. Momin MH
    . 2016. Evaluation of an immunochromatographic lateral flow assay (OXA-48 K-SeT) for rapid detection of OXA-48-like carbapenemases in Enterobacteriaceae. J Clin Microbiol54:471–473. doi:10.1128/JCM.02900-15.
    OpenUrlAbstract/FREE Full Text
  4. 4.↵
    1. Glupczynski Y,
    2. Evrard S,
    3. Ote I,
    4. et al
    . 2016. Evaluation of two new commercial immunochromatographic assays for the rapid detection of OXA-48 and KPC carbapenemases from cultured bacteria. J Antimicrob Chemother71:1217–1222. doi:10.1093/jac/dkv472.
    OpenUrlCrossRefPubMed
  5. 5.↵
    1. Dortet L,
    2. Jousset A,
    3. Sainte-Rose V,
    4. Cuzon G,
    5. Naas T
    . 2016. Prospective evaluation of the OXA-48 K-SeT assay, an immunochromatographic test for the rapid detection of OXA-48-type carbapenemases. J Antimicrob Chemother71:1834–1840. doi:10.1093/jac/dkw058.
    OpenUrlCrossRefPubMed
  6. 6.↵
    1. Meunier D,
    2. Vickers A,
    3. Pike R,
    4. Hill RL,
    5. Woodford N,
    6. Hopkins KL
    . 2016. Evaluation of the K-SeT R.E.S.I.S.T. immunochromatographic assay for the rapid detection of KPC and OXA-48-like carbapenemases. J Antimicrob Chemother71:2357–2359. doi:10.1093/jac/dkw113.
    OpenUrlCrossRefPubMed
  7. 7.↵
    1. Oikonomou O,
    2. Liakopoulos A,
    3. Phee LM,
    4. Betts J,
    5. Mevius D,
    6. Wareham DW
    . 2016. Providencia stuartii isolates from Greece: co-carriage of cephalosporin (blaSHV-5, blaVEB-1), carbapenem (blaVIM-1), and aminoglycoside (rmtB) resistance determinants by a multidrug-resistant outbreak clone. Microb Drug Resist22:379–386. doi:10.1089/mdr.2015.0215.
    OpenUrlCrossRef
  8. 8.↵
    1. Dallene C,
    2. Da Costa A,
    3. Decre D,
    4. Favier C,
    5. Arlet G
    . 2010. Development of a set of multiplex PCR assays for the detection of genes encoding β-lactamases in Enterobacteriaceae. J Antimicrob Chemother65:490–495. doi:10.1093/jac/dkp498.
    OpenUrlCrossRefPubMedWeb of Science
PreviousNext
Back to top
Download PDF
Citation Tools
Rapid Detection of Carbapenemases in Enterobacteriaceae: Evaluation of the Resist-3 O.K.N. (OXA-48, KPC, NDM) Lateral Flow Multiplexed Assay
David W. Wareham, Muhd Haziq F. Abdul Momin
Journal of Clinical Microbiology Mar 2017, 55 (4) 1223-1225; DOI: 10.1128/JCM.02471-16

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Print

Alerts
Sign In to Email Alerts with your Email Address
Email

Thank you for sharing this Journal of Clinical Microbiology article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Rapid Detection of Carbapenemases in Enterobacteriaceae: Evaluation of the Resist-3 O.K.N. (OXA-48, KPC, NDM) Lateral Flow Multiplexed Assay
(Your Name) has forwarded a page to you from Journal of Clinical Microbiology
(Your Name) thought you would be interested in this article in Journal of Clinical Microbiology.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Rapid Detection of Carbapenemases in Enterobacteriaceae: Evaluation of the Resist-3 O.K.N. (OXA-48, KPC, NDM) Lateral Flow Multiplexed Assay
David W. Wareham, Muhd Haziq F. Abdul Momin
Journal of Clinical Microbiology Mar 2017, 55 (4) 1223-1225; DOI: 10.1128/JCM.02471-16
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Top
  • Article
    • LETTER
    • ACKNOWLEDGMENTS
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

KEYWORDS

KPC
NDM
OXA-48
rapid diagnostics
carbapenems
Bacterial Proteins
bacteriological techniques
carbapenem-resistant Enterobacteriaceae
Chromatography, Affinity
Diagnostic Tests, Routine
Enterobacteriaceae Infections
beta-lactamases

Related Articles

Cited By...

About

  • About JCM
  • Editor in Chief
  • Board of Editors
  • Editor Conflicts of Interest
  • For Reviewers
  • For the Media
  • For Librarians
  • For Advertisers
  • Alerts
  • RSS
  • FAQ
  • Permissions
  • Journal Announcements

Authors

  • ASM Author Center
  • Submit a Manuscript
  • Article Types
  • Resources for Clinical Microbiologists
  • Ethics
  • Contact Us

Follow #JClinMicro

@ASMicrobiology

       

ASM Journals

ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology.

About ASM | Contact Us | Press Room

 

ASM is a member of

Scientific Society Publisher Alliance

 

American Society for Microbiology
1752 N St. NW
Washington, DC 20036
Phone: (202) 737-3600

 

Copyright © 2021 American Society for Microbiology | Privacy Policy | Website feedback

Print ISSN: 0095-1137; Online ISSN: 1098-660X