title: Matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification of nonfermenting gram-negative bacilli isolated from cystic fibrosis patients.
abstract: The identification of nonfermenting gram-negative bacilli isolated from cystic fibrosis (CF) patients is usually achieved by using phenotype-based techniques and eventually molecular tools. These techniques remain time-consuming, expensive, and technically demanding. We used a method based on matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) for the identification of these bacteria. A set of reference strains belonging to 58 species of clinically relevant nonfermenting gram-negative bacilli was used. To identify peaks discriminating between these various species, the profile of 10 isolated colonies obtained from 10 different passages was analyzed for each referenced strain. Conserved peaks with a relative intensity greater than 0.1 were retained. The spectra of 559 clinical isolates were then compared to that of each of the 58 reference strains as follows: 400 Pseudomonas aeruginosa, 54 Achromobacter xylosoxidans, 32 Stenotrophomonas maltophilia, 52 Burkholderia cepacia complex (BCC), 1 Burkholderia gladioli, 14 Ralstonia mannitolilytica, 2 Ralstonia pickettii, 1 Bordetella hinzii, 1 Inquilinus limosus, 1 Cupriavidus respiraculi, and 1 Burkholderia thailandensis. Using this database, 549 strains were correctly identified. Nine BCC strains and one R. mannnitolilytica strain were identified as belonging to the appropriate genus but not the correct species. We subsequently engineered BCC- and Ralstonia-specific databases using additional reference strains. Using these databases, correct identification for these species increased from 83 to 98% and from 94 to 100% of cases, respectively. Altogether, these data demonstrate that, in CF patients, MALDI-TOF-MS is a powerful tool for rapid identification of nonfermenting gram-negative bacilli.
authors.affiliation: Assistance Publique-Hôpitaux de Paris, Laboratoire de Microbiologie, Hôpital Necker-Enfants Malades, Paris, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Degand
authors.forename: Etienne
authors.initials: E
authors.lastname: Carbonnelle
authors.forename: Brunhilde
authors.initials: B
authors.lastname: Dauphin
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Beretti
authors.forename: Muriel
authors.initials: M
authors.lastname: Le Bourgeois
authors.forename: Isabelle
authors.initials: I
authors.lastname: Sermet-Gaudelus
authors.forename: Christine
authors.initials: C
authors.lastname: Segonds
authors.forename: Patrick
authors.initials: P
authors.lastname: Berche
authors.forename: Xavier
authors.initials: X
authors.lastname: Nassif
authors.forename: Agnès
authors.initials: A
authors.lastname: Ferroni
mesh_terms: D003550:Cystic Fibrosis / Q000150:complications*; D005285:Fermentation; D006090:Gram-Negative Bacteria / Q000145:classification* / Q000302:isolation & purification* / Q000378:metabolism; D016905:Gram-Negative Bacterial Infections / Q000382:microbiology*; D006801:Humans; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*