Carbapenemase-producing Pseudomonas aeruginosa from central Greece: molecular epidemiology and genetic analysis of class I integrons

BMC Infectious Diseases

Efthymia Petinaki⁴  Vivi Miriagou¹  Amalia D Karagouni⁵  Alexandros Theodosiou²  Efstathios A Katsifas⁵  Angeliki Mavroidi³  Apostolos Liakopoulos⁵ 

[1] Laboratory of Bacteriology, Hellenic Pasteur Institute, Athens, Greece;Department of Urology, University Hospital of Larissa, Larissa, Greece;Department of Microbiology, University Hospital of Larissa, Larissa, Greece;Department of Microbiology, Medical School, University of Thessaly, Biopolis, Larissa, Greece;Faculty of Biology, Microbiology Group, University of Athens, Athens, Greece

关键词: MLST;    Integrons;    VIM;    Carbapenems;    MDR;    Pseudomonas aeruginosa;   

Others  :  1145668
DOI  :  10.1186/1471-2334-13-505

 received in 2013-06-18, accepted in 2013-10-20,  发布年份 2013

【 摘 要 】

Background

Multidrug-resistant Pseudomonas aeruginosa is a serious challenge for antimicrobial therapy of nosocomial infections, as it possesses several mechanisms of antimicrobial resistance. In Central Greece, a sudden increase of infections caused by carbapenem-resistant P. aeruginosa was observed during 2011, indicating the need for further analysis.

Methods

Five-hundred and sixty-eight P. aeruginosa isolates were collected consecutively during an 8-month period in 2011 from inpatients treated in three hospitals in the Thessaly region (1,000,000 habitants) of Greece. Carbapenem-resistant P. aeruginosa (n = 284) were characterized by antimicrobial susceptibility testing and β-lactamase content, and the genetic relatedness of carbapenemase-producing isolates was assessed by BOX-PCR, multilocus sequence typing, and eBURST analysis. Mapping of the class I integrons of Verona integron-encoded metallo-β-lactamase (VIM)-carrying isolates was also performed, and clinical data of the VIM producers were reviewed.

Results

Eighty (14.1%) out of the 568 P. aeruginosa isolates recovered from clinical specimens were VIM producers. Multilocus sequence typing revealed high prevalence of the international clones ST111 and ST235 among blaVIM-2- and blaVIM-4-positive isolates, respectively. blaVIM-17 was identified in an isolate of a novel sequence type (ST1457). bla_VIM gene cassettes were carried by five distinct class I integrons, including two novel ones.

Conclusions

Since the first report of VIM-producing P. aeruginosa in 2000, this microorganism still remains among the most prevalent multidrug resistant pathogens in Greece. The spread of VIM-producers belonging to the most common international clones (ST111 and ST235), the spread of integrons of divergent structures, and the emergence of novel integrons underscore their ongoing evolution.

【 授权许可】

   
2013 Liakopoulos et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

【 参考文献 】

• [1]Tomás M, Doumith M, Warner M, Turton JF, Beceiro A, Bou G, Livermore DM, Woodford N: Efflux pumps, OprD porin, AmpC b -lactamase, and multiresistance inPseudomonas aeruginosaisolates from cystic fibrosis patients. Antimicrob Agents Chemother 2010, 54:2219-2224.
• [2]Weldhagen GF, Poirel L, Nordmann P: Ambler class A extended-spectrum β-lactamases in Pseudomonas aeruginosa: novel developments and clinical impact. Antimicrob Agents Chemother 2003, 47:2385-2392.
• [3]Queenan AM, Bush K: Carbapenamases: the versatile β-lactamases. Clin Microbiol Rev 2007, 20:440-458.
• [4]Sacha P, Wieczorek P, Hauschild T, Zórawski M, Olszańska D, Tryniszewska E: Metallo-beta-lactamases of pseudomonas aeruginosa–a novel mechanism resistance to beta-lactam antibiotics. Folia Histochem Cytobiol 2008, 46:137-142.
• [5]Tzouvelekis LS, Markogiannakis A, Psichogiou M, Tassios PT, Daikos GL: Carbapenemases in Klebsiella pneumoniae and other Enterobacteriaceae: an evolving crisis of global dimensions. Clin Microbiol Rev 2012, 25:682-707.
• [6]Lolans K, Queenan AM, Bush K, Sahud A, Quinn JP: First nosocomial outbreak of Pseudomonas aeruginosa producing-integron borne metallo-β-lactamase (VIM-2) in the United States. Antimicrob Agents Chemother 2005, 49:3538-3540.
• [7]Rossolini GM, Luzzaro F, Migliavacca R, Mugnaioli C, Pini B, Luca F, Perilli M, Pollini S, Spalla M, Amicosante G, Toniolo A, Pagani L: First countrywide survey of acquired metallo-β-lactamases in gram-negative pathogens in Italy. Antimicrob Agents Chemother 2008, 52:4023-4029.
• [8]Patzer JA, Walsh TR, Weeks J, Dzierżanowska D, Toleman MA: Emergence and persistence of integron structures harbouring VIM genes in the children’s memorial health institute, Warsaw, Poland, 1998–2006. J Antimicrob Chemother 2009, 63:269-273.
• [9]Lee K, Lim JB, Yum JH, Yong D, Chong Y, Kim JM, Livermore DM: blaVIM-2 cassette-containing novel integrons in metallo-β-lactamase-producing Pseudomonas aeruginosa and Pseudomonas putida isolates disseminated in a Korean hospital. Antimicrob Agents Chemother 2002, 46:1053-1058.
• [10]Zhao WH, Chen G, Ito R, Hu ZQ: Revelance of resistance levels to carbapenems and integron-borne blaIMP-1,blaIMP-7, blaIMP-10 and blaVIM-2 in clinical isolates of Pseudomonas aeruginosa. J Med Microbiol 2009, 58:1080-1085.
• [11]Viedma E, Juan C, Villa J, Barrado L, Orellana MA, Sanz F, Otero JR, Oliver A, Chaves F: VIM-2–producing multidrug-resistant Pseudomonas aeruginosa clone ST175, Spain. Emerg Infect Dis 2012, 18:1235-1241.
• [12]Woodford N, Turton JF, Livermore DM: Multiresistant gram-negative bacteria: the role of high-risk clones in the dissemination of antibiotic resistance. FEMS Microbiol Rev 2011, 35:736-755.
• [13]Giske CG, Libish B, Colinon C, Scoulica E, Pagani L, Füzi M, Kronvall G, Rossolini GM: Establishing clonal relationships between VIM-1-like metallo-β-lactamase-producing Pseudomonas aeruginosa strains from four European countries by multilocus sequence typing. J Clin Microbiol 2006, 44:4309-4315.
• [14]Curran B, Jonas D, Grundmann H, Pitt T, Dowson CG: Development of multilocus sequence typing scheme for the opportunistic pathogen Pseudomonas aeruginosa. Clin Microbiol 2004, 42:5644-5649.
• [15]Clinical and Laboratory Standards Institute: Performance standards for antimicrobial susceptibility testing: twenty-first informational supplement, M100-S21. Wayne, PA, USA: Clinical and Laboratory Standards Institute (CLSI); 2011.
• [16]Ellington MJ, Kistler J, Livermore DM, Woodford N: Multiplex PCR for rapid detection of genes encoding acquired metallo-β-lactamases. J Antimicrob Chemother 2007, 59:321-322.
• [17]Marques ASA, Marchaison A, Gardan L, Samson R: BOX-PCR-based identification of bacterial species belonging to Pseudomonas syringae-P. viridiflava group. Genet Mol Biol 2008, 31:106-115.
• [18]Feil EJ, Li BC, Aanensen DM, Hanage WP, Spratt BG: eBURST: inferring patterns of evolutionary descent among clusters of related bacterial genotypes from multilocus sequence typing data. J Bacteriol 2004, 186:1518-1530.
• [19]Samuelsen Ø, Toleman MA, Sundsfjord A, Rydberg J, Leegaard TM, Walder M, Lia A, Ranheim TE, Rajendra Y, Hermansen NO, Walsh TR, Giske CG: Molecular epidemiology of metallo-β-lactamase-producing Pseudomonas aeruginosa isolates from Norway and Sweden shows import of international clones and local clonal expansion. Antimicrob Agents Chemother 2010, 54:346-352.
• [20]Siarkou VI, Vitti D, Protonotariou E, Ikonomidis A, Sofianou D: Molecular epidemiology of outbreak-related Pseudomonas aeruginosa strains carrying the novel variant blaVIM-17 metallo-β-lactamase gene. Antimicrob Agents Chemother 2009, 53:1325-1330.
• [21]Koutsogiannou M, Drougka E, Liakopoulos A, Jelastopulu E, Petinaki E, Anastassiou ED, Spiliopoulou I, Christofidou M: Spread of multidrug resistant Pseudomonas aeruginosa clones in a university hospital. J Clin Microbiol 2013, 51:665-668.
• [22]Hentschke M, Goritzkaa V, Camposa CB, Merkelb P, Ilchmannc C, Lellekd H, Scherpea S, Aepfelbachera M, Rohdea H: Emergence of carbapenemases in gram-negative bacteria in Hamburg, Germany. Diagn Microbiol Infect Dis 2011, 71:312-315.
• [23]Larche J, Pouillot F, Essoh C, Libisch B, Straut M, Lee JC, Soler C, Lamarca R, Gleize E, Gabard J, Vergnaud G, Pourcel C: Rapid identification of international multidrug-resistant Pseudomonas aeruginosa clones by multiple-locus variable number of tandem repeats analysis and investigation of their susceptibility to lytic bacteriophages. Antimicrob Agents Chemother 2012, 56:6175-6180.
• [24]Cabot G, Ocampo-Sosa AA, Dominguez MA, Gago JF, Juan C, Tubau F, Rodriguez C, Moya B, Pena C, Martinez-Martinez L, Oliver A: Genetic markers of widespread extensively drug-resistant Pseudomonas aeruginosa high-risk clones. Antimicrob Agents Chemother 2012, 56:6349-6357.
• [25]Vatcheva-Dobrevska R, Mulet X, Ivanov I, Zamorano L, Dobreva E, Velinov T, Kantardjiev T, Oliver A: Molecular epidemiology and multidrug resistance mechanisms of Pseudomonas aeruginosa isolates from Bulgarian hospitals. Microb Drug Resist 2013, 19(5):335-361. Epub ahead of print
• [26]Pollini S, Antonelli A, Venturelli C, Maradei S, Veggetti A, Bracco S, Rumpianesi F, Luzzaro F, Rossolini GM: Acquisition of plasmid-borne blaIMP-19 gene by a VIM-1-positive Pseudomonas aeruginosa of the sequence type 235 epidemic lineage. J Antimicrob Chemother 2013, 68:722-724.
• [27]Park YS, Lee H, Chin BS, Han SH, Hong SG, Hong SK, Kim HY, Uhg Y, Shin HB, Choo EJ, Han S-H, Song W, Jeong SH, Lee K, Kim JM: Acquisition of extensive drug-resistant Pseudomonas aeruginosa among hospitalized patients: risk factors and resistance mechanisms to carbapenems. J Hosp Infect 2011, 79:54-58.
• [28]Tsakris A, Pournaras S, Woodford N, Palepou MF, Babini GS, Douboyas J, Livermore DM: Outbreak of infections caused by Pseudomonas aeruginosa producing VIM-1 carbapenemase in Greece. J Clin Microbiol 2000, 38:1290-1292.
• [29]Mavroidi A, Tsakris A, Tzelepi E, Pournaras S, Loukova V, Tzouvelekis LS: Carbapenem-hydrolysing VIM-2 metallo- beta-lactamase in Pseudomonas aeruginosa from Greece. J Antimicrob Chemother 2000, 46:1041-1042.
• [30]Pournaras S, Maniati M, Petinaki E, Tzouvelekis LS, Tsakris A, Legakis NJ, Maniatis AN: Hospital outbreak of multiple clones of Pseudomonas aeruginosa carrying the unrelated metallo-β-lactamase gene variants blaVIM-2 and blaVIM-4. J Antimicrob Chemother 2003, 51:1409-1414.