Antimicrobial Susceptibility Studies Molecular epidemiology characterization of OXA-23 carbapenemase-producing Acinetobacter baumannii isolated from 8 Brazilian hospitals using repetitive sequence–based PCR ☆ Juliette M. Cieslinski a , Lavinia Arend b , Felipe F. Tuon c, ⁎, Ethianne P. Silva a , Rafael G.S. Ekermann a , Libera Maria Dalla-Costa d, e , Paul G. Higgins f , Harald Seifert f , Marcelo Pilonetto a, b, ⁎ a Department of Microbiology, School of Health and Biosciences, Pontifícia Universidade Católica do Paraná, Curitiba, PR, Brazil b Bacteriology Section, Laboratório Central de Saúde Pública do Estado LACEN-PR, São José dos Pinhais, PR, Brazil c Division of Infectious Diseases, Universidade Federal do Parana, Curitiba, PR, Brazil d Hospital de Clinicas, Universidade Federal do Parana, Curitiba, PR, Brazil e Faculdades e Instituto de Pesquisa Pele Pequeno Principe, Curitiba, PR Brazil f Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, Cologne, Germany abstract article info Article history: Received 13 January 2013 Received in revised form 27 July 2013 Accepted 31 July 2013 Available online 24 September 2013 Keywords: Acinetobacter baumannii Automated repetitive-sequence-based PCR Molecular epidemiology DiversiLab The typing of multidrug-resistant Acinetobacter baumannii isolates is important for the control and prevention of hospital outbreaks. This study aimed to analyze the molecular epidemiology of 46 OXA-23 carbapenemase- producing A. baumannii strains and compare them to previously described local and international clones (ICs). Isolates were recovered during May 2009–August 2011, from 8 different hospitals in the state of Parana (Brazil). The molecular profiles were determined by repetitive extragenic palindromic PCR. Seven different clusters were identified (A to G). Thirty-two isolates were clustered in the same pattern (clone A), which belong to IC 4. © 2013 Elsevier Inc. All rights reserved. 1. Brief report Acinetobacter baumannii is a Gram-negative hospital-acquired pathogen. The analysis of the molecular epidemiology of nosocomial infections caused by carbapenem-resistant A. baumannii (CRAb) has been performed to help develop efficient guidelines to control its spread (Koeleman et al., 2001; Toledo et al., 2012). Epidemiological tools may be used to investigate outbreaks, to confirm and define the transmission behavior of 1 or more clones, to test hypotheses regarding the clonal origin or transmission modes, and to monitor and control epidemic reservoirs. Repetitive extragenic palindromic PCR (rep-PCR) is considered reliable, reproducible, fast, and discriminatory (Perez et al., 2010). This method is based on the amplification of repetitive sequences of highly conserved DNA that are present in multiple copies at different positions in the genome of bacteria. DiversiLab is a semi-automated rep-PCR– based system that has been developed for such investigations. In the current study, we analyzed the molecular epidemiological features of 46 CRAb strains isolated during 2009–2011 from 8 hospitals in southern Brazil (Paraná State—total area: 199,314 km 2 ; approximately 10 million inhabitants), by using rep-PCR with DiversiLab®. Also, we included in our study representative strains from a previous study in the same region (Schimith Bier et al., 2010) and reference strains corresponding to the 8 international clones (ICs) (Higgins et al., 2010). The strains were isolated from blood (n = 29), catheters (n = 14), peritoneal fluid (n = 2), and cerebrospinal fluid (n = 1). A. baumannii was initially identified by Vitek 2 (bioMérieux®, Marcy-LÉtoile, France) and confirmed by gyrB multiplex PCR (Higgins et al., 2007). Susceptibility testing was performed by the disk diffu- sion (CLSI, 2013). PCR analysis was performed to check for the pres- ence of bla OXA-23 (Correa et al., 2012). DNA extraction, amplification with DiversiLab® Acinetobacter DNA Fingerprinting Kit was previously described (Carretto et al., 2008). Isolates that showed N95% clustering were considered related (Saeed et al., 2006). Our PCR results showed that all 46 isolates (100%) carried the bla(OXA-23) gene. The first isolate of A. baumannii with the enzyme Diagnostic Microbiology and Infectious Disease 77 (2013) 337–340 ☆ Other authors: The authors are not part of any associations or commercial relationships that might represent conflicts of interest in the writing of this study (e.g., pharmaceutical stock ownership, consultancy, advisory board membership, relevant patents, or research funding). ⁎ Corresponding author. Tel.: +55-41-3271-2169. E-mail address: m.pilonetto@pucpr.br (M. Pilonetto). 0732-8893/$ – see front matter © 2013 Elsevier Inc. All rights reserved. http://dx.doi.org/10.1016/j.diagmicrobio.2013.07.018 Contents lists available at ScienceDirect Diagnostic Microbiology and Infectious Disease journal homepage: www.elsevier.com/locate/diagmicrobio