title: Rapid species identification and epidemiological analysis of carbapenem-resistant Acinetobacter spp. by a PCR-based open reading frame typing method.
abstract: The spread of carbapenem-resistant Acinetobacter spp. has become a global problem. In this study, 18 carbapenem-resistant Acinetobacter calcoaceticus-baumannii (ACB) complexes, identified using a conventional biochemical method at our hospital during 2004-2013, were studied for species identification and epidemiological analyses. Species identification was performed using matrix-assisted laser desorption ionization-time-of-flight MS, a partial sequence analysis of rpoB and a PCR-based ORF typing (POT) method. The POT method can not only identify the species of ACB complexes but also simultaneously determine the international epidemic clones and the genetic identities of Acinetobacterbaumannii in several hours. Carbapenem resistance gene detection by PCR, molecular epidemiological analysis by PFGE and Pasteur Institute multilocus sequence typing (MLST) analysis were performed. All three methods identified 18 isolates as A. baumannii (n=10), Acinetobacterpittii (n=4) and Acinetobacternosocomialis (n=4). A metallo-β-lactamase gene in all strains of A. pittii and A. nosocomialis and an ISAba1 gene in the upstream of the blaOXA-51-like gene in eight strains of A. baumannii were detected, respectively, as carbapenemase-related genes. Results from PFGE demonstrated that nine strains of A. baumannii were closely related genetically. Results of MLST analysis showed that A. baumannii are classifiable to sequence type 2. These results were consistent with those obtained using the POT method. This POT method can easily and rapidly identify the international epidemic clones and the identities of A. baumannii. It can be a useful tool for infection control.
authors.affiliation: Division of Central Clinical Laboratory, Iwate Medical University Hospital, Morioka, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Yamada
authors.affiliation: Division of Central Clinical Laboratory, Iwate Medical University Hospital, Morioka, Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Endo
authors.affiliation: Division of Central Clinical Laboratory, Iwate Medical University Hospital, Morioka, Japan.
authors.forename: Kaori
authors.initials: K
authors.lastname: Sawase
authors.affiliation: Division of Central Clinical Laboratory, Iwate Medical University Hospital, Morioka, Japan.
authors.forename: Marie
authors.initials: M
authors.lastname: Anetai
authors.affiliation: Division of Central Clinical Laboratory, Iwate Medical University Hospital, Morioka, Japan.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Narita
authors.affiliation: Division of Central Clinical Laboratory, Iwate Medical University Hospital, Morioka, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Hatakeyama
authors.affiliation: Division of Central Clinical Laboratory, Iwate Medical University Hospital, Morioka, Japan.
authors.forename: Katsunori
authors.initials: K
authors.lastname: Ishifuji
authors.affiliation: Division of Central Clinical Laboratory, Iwate Medical University Hospital, Morioka, Japan.
authors.forename: Makiko
authors.initials: M
authors.lastname: Kurota
authors.affiliation: Department of Laboratory Medicine, Iwate Medical University School of Medicine, Morioka, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Suwabe
mesh_terms: D000150:Acinetobacter / Q000145:classification* / Q000187:drug effects / Q000302:isolation & purification; D000151:Acinetobacter Infections / Q000175:diagnosis* / Q000382:microbiology*; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics; D015780:Carbapenems / Q000494:pharmacology; D016000:Cluster Analysis; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D005838:Genotype; D006801:Humans; D058889:Molecular Typing / Q000379:methods*; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013997:Time Factors; D018440:beta-Lactam Resistance; D001618:beta-Lactamases / Q000235:genetics
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D015780:Carbapenems; D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit; D001618:beta-Lactamases; C063912:carbapenemase