Investigation of Metallo Beta Lactamases and Oxacilinases in Carbapenem Resistant Acinetobacter baumannii Strains Isolated from Inpatients

Abstract

Background: Resistance to beta-lactam antibiotics is widespread among Acinetobacter strains. Plasmid-mediated metallo beta lactamases (MBL) are responsible for carbapenem resistance, as are oxacillinases (OXA). In recent years, MBL producing carbapenem-resistant strains have been reported in the world and in Turkey in increasing rates. In our country, besides the OXA 51- like enzyme which is inherent in A. baumannii strains, OXA 58-like and OXA 23-like carbapenemases producing strains have also been widely detected. In addition, Verona Imipenemase (VIM) and (IMP)-type MBL have been reported in some centers. Aims: The aim of our study was to investigate the presence of carbapenemases in Acinetobacter strains isolated from hospitalized patients in Edirne. Study Design: Cross-sectional study. Methods: A total of 52 imipenem-resistant A. baumannii strains isolated between January and March 2013 were investigated. The presence of MBL was described phenotypically by the combined disk diffusion test (CDDT), double disk synergy test (DDST), MBL Etest (only performed in 28 strains) and modified Hodge test. blaIMP, blaVIM, blaGIM, blaSIM, blaSPM genes and blaOXA-23, blaOXA-51, blaOXA-40, blaOXA-58 genes were investigated by multiplex polymerase chain reaction (PCR). The blaNDM-1 gene was determined by PCR. Results: By modified Hodge test, 50 strains (96%) were found to be MBL positive. Positivity of MBL was 21% by both CDDT (0.1 M EDTA) and DDST. Twenty-four of 28 strains (85.7%) were positive by MBL E-test. OXA 23-like and OXA 51-like carbapenemases were detected in all strains, but OXA 58-like and OXA 40-like carbapenemases-producing A. baumannii were not detected. Also, MBL genes were not detected by genotypic methods. Conclusion: Only OXA 23-like carbapenemase was responsible for carbapenem resistance in carbapenemresistant Acinetobacter strains in Edirne. The MBLproducing Acinetobacter strain is not yet a problem in our hospital. MBL resistance was found by phenotyping tests, which must be confirmed by genotypic methods; multiplex PCR tests can be easily used for screening MBL

Keywords

• Acinetobacter baumannii, metallo beta lactamases, oxacillinases

References

1. 1. Giamarellou H, Antoniadou A, Kanellakopoulou K. Acinetobacter baumannii: a universal threat to public health? Int J Antimicrob Agents 2008;32:106-19. [CrossRef]
2. 2. Cornaglia G, Giamarellou H, Rossolini GM. Metallo-βlactamases: a last frontier for β-lactams? Lancet Infect Dis 2011;11:381-93. [CrossRef]
3. 3. Gur D, Korten V, Unal S, Deshpande LM, Castanheira M. Increasing carbapenem resistance due to the clonal dissemination of oxacillinase (OXA-23 and OXA-58)-producing Acinetobacter baumannii: report from the Turkish SENTRY Program sites. J Med Microbiol 2008;57:1529-32. [CrossRef]
4. 4. Vahaboglu H, Budak F, Kasap M, Gacar G, Torol S, Karadenizli A, et al. High prevalence of OXA-51-type class D betalactamases among ceftazidime-resistant clinical isolates of Acinetobacter spp.: co-existence with OXA-58 in multiple centres. J Antimicrob Chemother 2006;58:537-42. [CrossRef]
5. 5. Sarı AN, Biçmen M, Gülay Z. The First Report on the Outbreak of OXA-24/40-Like Carbapenemase-Producing Acinetobacter baumannii in Turkey. Jpn J Infect Dis 2013;66:439-42.
6. 6. Zarrilli R, Pournaras S, Giannouli M, Tsakris A. Global evolution of multidrug resistant Acinetobacter baumannii clonal lineages. Int J Antimicrob Agents 2013;41:11-9. [CrossRef]
7. 7. Bahar G, Mazzariol R, Koncan R, Mert A, Fontana R, Rossolini GM, et al. Detection of VIM-5 metallo-betalactamase in a Pseudomonas aeruginosa clinical isolate from Turkey. J Antimicrob Chemother 2004;54:282-3. [CrossRef]
8. 8. Ozgumus OB, Caylan R, Tosun I, Sandalli C, Aydin K, Koksal I. Molecular epidemiology of clinical Pseudomonas aeruginosa isolates carrying IMP-1 metallo-beta-lactamase gene in a University Hospital in Turkey. Microb Drug Resist 2007;13:191-8. [CrossRef]

9. 9. Yakupogullari Y, Poirel L, Bernabeu S, Kizirgil A, Nordmann P. Multidrug-resistant Pseudomonas aeruginosa isolate co-expressing extended- spectrum beta-lactamase PER-1 and metallo-beta-lactamase VIM2 from Turkey. J Antimicrob Chemother 2008;61:221-2. [CrossRef]
10. 10. Alp E, Perçin D,Colakoğlu S, Durmaz S, KürkcüCA, Ekincioğlu P, et al. Moleculacharacterization of carbapenem-resistant Klebsiella pneumoniae in a tertiary university hospital in Turkey. J Hosp Infect 2013;84:178-80. [CrossRef]
11. 11. Aktaş Z, Kayacan CB. Investigation of metallo-beta-lactamase producing strains of Pseudomonas aeruginosa and Acinetobacter baumannii by E-test, disk synergy and PCR. Scand J Infect Dis 2008;40:320-5. [CrossRef]
12. 12. Lee K, Chong Y, Shin HB, Kim YA, Yong D, Yum JH. Modified Hodge and EDTA-disk synergy tests to screenmetallo-βlactamase-producing strains of Pseudomonas and Acinetobacter species. Clin Microbiol Infect 2001;7:88-91. [CrossRef]
13. 13. Franco MR, Caiaffa-Filho HH, Burattini MN, Rossi F. Metallobetalactamases among imipenem-resistant Pseudomonas aeruginosa in a Brazilian university hospital. Clinics (Sao Paulo) 2010;65:825-9. [CrossRef]
14. 14. Ellington MJ, Kistler J, Livermore DM, Woodford N. Multiplex PCR for rapid detection of genes encoding acquired metallo-betalactamases. J Antimicrob Chemother 2007;59:321-2. [CrossRef]
15. 15. Woodford N. Rapid Characterization of β lactamases by Multiplex PCR. In: Gillespie SH. and McHugh TD, editors. Antibiotic Resistance Protocols. 2th ed. London: Humana Press; 2010. p.181-92. [CrossRef]
16. 16. Zarfel G, Hoenigl M, Leitner E, Salzer HJ, Feierl G, Masoud L, et al. Emergence of New Delhi metallo-β-lactamase, Austria. Emerg Infect Dis 2011;17:129-30. [CrossRef]
17. 17. Esen S, Leblebicioglu H. Prevalence of nosocomial infections at intensive care units in Turkey: a multicentre 1-day point prevalence study. Scand J Infect Dis 2004;36:144-8. [CrossRef]
18. 18. Ayres HM, Furr JR, Russell AD. Effect of permeabilizers on antibiotic sensitivity of Pseudomonas aeruginosa. Lett Appl Microbiol 1999;28:13-6. [CrossRef]
19. 19. Denny B, West P, Panigrahi D. Effects of permeabilizers on antimicrobial susceptibility of Stenotrophomonas maltophilia and Acinetobacter spp. J Microbiol Immunol Infect 2003;36:72-6.
20. 20. Ulusoy Al M, Mumcuoğlu İ, Aksu N, Dolapçı İ, Karahan ZC, Baran I, ve ark. İmipenem dirençli Acinetobacter İzolatlarında metallo-beta-laktamaz üretiminin fenotipik ve genotipik yöntemlerle araştırılması. Türk Mikrobiyol Cem Derg 2011;41:29-36.
21. 21. Eser ÖK, Ergin A, Hasçelik G. Erişkin hastalardan izole edilen Acinetobacter türlerinde antimikrobiyal direnç ve metallo-betalaktamaz varlığı. Mikrobiyol Bul 2009;43:383-90.
22. 22. Türk Dağı H, Kuş H, Keyik Ş, Arslan U, Tuncer İ, Fındık D. Karbapenemlere Dirençli Acinetobacter baumannii Suşlarında Metallo-Beta-Laktamaz Varlığının Araştırılması. ANKEM Derg 2012;26:187-92.
23. 23. Rodrigues AC, Chang MR, Nóbrega GD, Rodrigues MS, Carvalho NC, Gomes BG, et al. Metallo-β-lactamase and genetic diversity of Pseudomonas aeruginosa in intensive care units in Campo Grande, MS, Brazil. Braz J Infect Dis 2011;15:195-9. [CrossRef]
24. 24. Ratkai C, Quinteira S, Grosso F, Monteiro N, Nagy E, Peixe L. Controlling for false positives: interpreting MBL Etest and MBL combined disc test for the detection of metallo-beta-lactamases. J Antimicrob Chemother 2009;64:657-8. [CrossRef]
25. 25. Chu YW, Cheung TK, Ngan JY, Kam KM. EDTA susceptibility leading to false detection of metallo-b-lactamase in Pseudomonas aeruginosa by E-test and an imipenem-EDTA disk method. Int J Antimicrob Agents 2005;26:340-1. [CrossRef]
26. 26. Kulah C, Mooij MJ, Comert F, Aktas E, Celebi G, Ozlu N, et al. Characterisation of carbapenem-resistant Acinetobacter baumannii outbreak strains producing OXA-58 in Turkey. Int J Antimicrob Agents 2010;36:114-8. [CrossRef]
27. 27. Ozen N, Ergani A, Naas T, Ogunc D, Gultekin M, Colak D, et al. Outbreak of carbapenem-resistant Acinetobacter baumannii producing the carbapenemase OXA-58 in Turkey. Open Antimicrob Agents J 2009;1:1-8. [CrossRef]
28. 28. Pournaras S, Markogiannakis A, Ikonomidis A, Kondyli L, Bethimouti K, Maniatis A, et al. Outbreak of multiple clones of imipenem-resistant Acinetobacter baumannii isolates expressing OXA-58 carbapenemase in an intensive care unit. J Antimicrob Chemother 2006;57:557-61. [CrossRef]
29. 29. Liakopoulos A, Miriagou V, Katsifas EA, Karagouni AD, Daikos GL, Tzouvelekis LS, et al. Identification of OXA-23-producing Acinetobacter baumannii in Greece, 2010 to 2011. Euro Surveill 2012;17.
30. 30. Strateva T, Markova B, Marteva-Proevska Y, Ivanova D, Mitov I. Widespread dissemination of multidrug-resistant Acinetobacter baumannii producing OXA-23 carbapenemase and ArmA 16S ribosomal RNA methylase in a Bulgarian university hospital. Braz J Infect Dis 2012;16:307-10. [CrossRef