title: Development and Validation of a Lateral Flow Immunoassay for Rapid Detection of NDM-Producing Enterobacteriaceae.
abstract: The global spread of carbapenemase-producingEnterobacteriaceae(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-producingEnterobacteriaceaefrom 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-producingEnterobacteriaceae.
authors.affiliation: Service de Pharmacologie et Immunoanalyse, CEA, INRA, Université Paris-Saclay, Gif-sur-Yvette, France.
authors.forename: Hervé
authors.initials: H
authors.lastname: Boutal
authors.affiliation: EA7361, Université Paris-Sud, Université Paris-Saclay, LabEx Lermit, Bacteriology-Hygiene Unit, APHP, Hôpital Bicêtre, Le Kremlin-Bicêtre, France.|Associated French National Reference Center for Antibiotic Resistance: Carbapenemase-Producing Enterobacteriaceae, Le Kremlin-Bicêtre, France.
authors.forename: Thierry
authors.initials: T
authors.lastname: Naas
authors.affiliation: Service de Pharmacologie et Immunoanalyse, CEA, INRA, Université Paris-Saclay, Gif-sur-Yvette, France.
authors.forename: Karine
authors.initials: K
authors.lastname: Devilliers
authors.affiliation: EA7361, Université Paris-Sud, Université Paris-Saclay, LabEx Lermit, Bacteriology-Hygiene Unit, APHP, Hôpital Bicêtre, Le Kremlin-Bicêtre, France.
authors.forename: Saoussen
authors.initials: S
authors.lastname: Oueslati
authors.affiliation: EA7361, Université Paris-Sud, Université Paris-Saclay, LabEx Lermit, Bacteriology-Hygiene Unit, APHP, Hôpital Bicêtre, Le Kremlin-Bicêtre, France.|Associated French National Reference Center for Antibiotic Resistance: Carbapenemase-Producing Enterobacteriaceae, Le Kremlin-Bicêtre, France.
authors.forename: Laurent
authors.identifier: 0000-0001-6596-7384
authors.initials: L
authors.lastname: Dortet
authors.affiliation: EA7361, Université Paris-Sud, Université Paris-Saclay, LabEx Lermit, Bacteriology-Hygiene Unit, APHP, Hôpital Bicêtre, Le Kremlin-Bicêtre, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Bernabeu
authors.affiliation: EA7361, Université Paris-Sud, Université Paris-Saclay, LabEx Lermit, Bacteriology-Hygiene Unit, APHP, Hôpital Bicêtre, Le Kremlin-Bicêtre, France.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Simon
authors.affiliation: EA7361, Université Paris-Sud, Université Paris-Saclay, LabEx Lermit, Bacteriology-Hygiene Unit, APHP, Hôpital Bicêtre, Le Kremlin-Bicêtre, France herve.volland@cea.fr.
authors.forename: Hervé
authors.initials: H
authors.lastname: Volland
mesh_terms: D000073182:Carbapenem-Resistant Enterobacteriaceae / Q000201:enzymology / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D004756:Enterobacteriaceae Infections / Q000382:microbiology; D006801:Humans; D012680:Sensitivity and Specificity; D013997:Time Factors; D001618:beta-Lactamases / Q000032:analysis*
chemical_list: D001618:beta-Lactamases; C552864:beta-lactamase NDM-1
keywords: NDM-1; detection; lateral flow immunoassay