title: Real-time PCR assay allows detection of the New Delhi metallo-β-lactamase (NDM-1)-encoding gene in France.
abstract: In this study, we report the development of a rapid real-time polymerase chain reaction (PCR) assay with TaqMan probe to detect the New Delhi metallo-β-lactamase (NDM-1)-encoding gene directly from bacterial isolates. The specificity of the assay was verified in silico as well as with a large panel of 84 clinically relevant bacteria, including the Klebsiella pneumoniae NCTC 13443 NDM-1-positive reference strain. Using this assay retrospectively on a local series of 44 K. pneumoniae isolates from Marseille Hospitals (France), it was possible to detect and identify an NDM-1-producing K. pneumoniae strain isolated from bronchoalveolar lavage in April 2010 from a French patient repatriated from India after a motorbike accident. Standard PCR amplification and sequencing of the entire NDM-1 gene from this isolate was also performed and the amino acid sequence showed 100% homology with the NDM-1 protein from the K. pneumoniae reference strain. We believe that this real-time PCR assay would be a powerful tool that could be added to other molecular detection assays such as detection of KPC- or OXA-encoding genes for rapid screening and/or identification of carbapenem-resistant bacterial isolates from patients returning from the Asian continent that could be implemented in a point-of-care strategy.
authors.affiliation: Unité de Recherche sur Maladies Infectieuses et Tropicales Emergents, CNRS-IRD, UMR 6236, Faculté de Médecine et de Pharmacie, Université de Méditerranée Aix-Marseille II, 27 Bd Jean Moulin, 13385 Marseille Cedex 05, France.
authors.forename: Seydina M
authors.initials: SM
authors.lastname: Diene
authors.forename: Nicolas
authors.initials: N
authors.lastname: Bruder
authors.forename: Didier
authors.initials: D
authors.lastname: Raoult
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Rolain
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods*; D005602:France; D006801:Humans; D007710:Klebsiella Infections / Q000382:microbiology; D007711:Klebsiella pneumoniae / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D001618:beta-Lactamases / Q000235:genetics*
chemical_list: D001618:beta-Lactamases; C552864:beta-lactamase NDM-1