Advances in Clinical and Experimental Medicine
2019, vol. 28, nr 2, February, p. 249–254
Publication type: original article
Emergence of Enterobacteriaceae co-producing CTX-M-15, ArmA and PMQR in Poland
1 Department of Bacteriology, National Institute of Public Health–National Institute of Hygiene, Warszawa, Poland
Background. Plasmid-mediated extended-spectrum β-lactamases (ESBLs), 16S rRNA methylases and quinolone resistance mechanisms (PMQRs) are well-known agents conferring resistance to more than 1 antimicrobial in its group. The accumulation of these agents poses, therefore, a serious risk to public health.
Objectives. The objective of this study was to investigate the presence of common ß-lactamases and 16S rRNA methylases in Qnr-producing Enterobacteriaceae and their genetic relatedness.
Material and Methods. We examined 18 Qnr-producing isolates (Klebsiella pneumoniae n = 8, Enterobacter cloacae n = 6 and Escherichia coli n = 4) selected from a collection of 215 ciprofloxacin-resistant strains obtained from patients in a 1030-bed tertiary hospital from 1 March to 31 August 2010. The antibiotics minimum inhibitory concentration (MIC) was determined by E-test. The detection of common ß-lactamases, 16S rRNA methyltransferases and PMQR genes was performed by polymerase chain reaction (PCR) and sequencing. Genetic relatedness was assessed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST).
Results. All the isolates tested were susceptible to carbapenems and colistin, while 16 were multidrug-resistant. Thirteen, 2 and 2 isolates carried qnrB1, qnrA1 and qnrS1, respectively. Ten of 13 qnrB1-positive Enterobacteriaceae also carried genes encoding for aac(6’)-Ib-cr and at least 1 ESBL. The blaCTX-M-15 gene was the most common ESBL. The most prevalent combination of genes was qnrB1+aac(6’)-Ib-cr+blaTEM-1+blaCTX-M-15. Two isolates of K. pneumoniae and E. cloacae were found to bear multiple extended range resistance traits: ArmA, CTX-M-15, QnrB1, and AAC (6’)-Ib-cr. The PFGE showed that most of the isolates exhibited individual DNA patterns, whilst MLST assigned K. pneumoniae (n = 8) to 5 sequence types (STs) (ST15, ST323, ST336, ST147, and ST525), E. coli (n = 4) to 2 (ST131 and ST1431) and E. cloacae (n = 5) to 4 (ST90, ST89, ST133, and the novel ST407).
Conclusion. Our findings reveal the accumulation of resistance traits and their important role in spreading of multiresistant bacteria among hospitalized patients.
PMQR, ESBLs, MDR, Enterobacteriaceae, Poland
- European Centre for Disease Prevention and Control. Antimicrobial resistance surveillance in Europe 2013. Annual Report of the European Antimicrobial Resistance Surveillance Network (EARS-Net). Stockholm: ECDC , Sweden; 2014. http://ecdc.europa.eu/en/publications/Publications/antimicrobial-resistance-europe-2013.pdf.
- Hooper DC, Jacoby GA. Mechanisms of drug resistance: Quinolone resistance. Ann N Y Acad Sci. 2015;1354:13–31.
- Piekarska K, Wołkowicz T, Zacharczuk K, et al. Co-existence of plasmid-mediated quinolone resistance determinants and mutations in gyrA and parC among fluoroquinolone-resistant clinical Enterobacteriaceae isolated in tertiary hospital in Warsaw, Poland. Int J Antimicrob Agents. 2015;45(3):238–243.
- Dolejska M, Villa L, Dobiasova H, Fortini D, Feudi C, Carattoli A. Plasmid content of a clinically relevant Klebsiella pneumoniae clone from the Czech Republic producing CTX-M-15 and QnrB1. Antimicrob Agents Chemother. 2013;57(2):1073–1076.
- Filippa N, Carricajo A, Grattard F, et al. Outbreak of multidrug-resistant Klebsiella pneumoniae carrying qnrB1 and blaCTX-M-15 in a French intensive care unit. Ann Intensive Care. 2013;3:18.
- Tóth Á, Kocsis B, Damjanova I, et al. Fitness cost associated with resistance to fluoroquinolones is diverse across clones of Klebsiella pneumoniae and may select for CTX-M-15 type extended-spectrum ß-lactamase. Eur J Clin Microbiol Infect Dis. 2014;33(5):837–843.
- Luo Y, Yang J, Zhang Y, Ye L, Wang L, Guo L. Prevalence of β-lactamases and 16S rRNA methylase genes amongst clinical Klebsiella pneumoniae isolates carrying plasmid-mediated quinolone resistance determinants. Int J Antimicrob Agents. 2011;37(4):352–355.
- Zacharczuk K, Piekarska K, Szych J, et al. Plazmid-borne 16S rRNA methylase ArmA in aminoglicoside-resistant Klebsiella pneumoniae in Poland. J Med Microbiol. 2011;60(Pt 9):1306–1311.
- Pérez-Pérez FJ, Hanson ND. Detection of plasmid-mediated AmpC beta-lactamase genes in clinical isolates by using multiplex PCR. J Clin Microbiol. 2002;40(6):2153–2162.
- Piekarska K, Zacharczuk K, Wołkowicz T, et al. Distribution of 16S rRNA methylases among different species of aminoglycoside-resistant Enterobacteriaceae in tertiary care hospital in Poland. Adv Clin Exp Med. 2016;25(3):539–544.
- Miyoshi-Akiyama T, Hayakawa K, Ohmagari N, Shimojima M, Kirikae T. Multilocus Sequence Typing (MLST) for characterization of Enterobacter cloacae. PLoS One. 2013;8(6):e66358.
- Mellmann A, Bielaszewska M, Kock R, et al. Analysis of collection of hemolytic uremic syndrome-associated enterohemorrhagic Escherichia coli. Emerg Infect Dis. 2008;14(8):1287–1290.
- Diancourt L, Passet V, Verhoef J, Grimont PAD, Brisse S. Multilocus sequence typing of Klebsiella pneumoniae nosocomial isolates. J Clin Microbiol. 2005;43(8):4178–4182.
- Zacharczuk K, Piekarska K, Szych J, et al. Emergence of Klebsiella pneumoniae coproducing KPC-2 and 16S rRNA methylase ArmA in Poland. Antimicrob Agents Chemother. 2011;55(1):443–446.
- Briales A, Rodríguez-Martínez JM, Velasco C, et al. Prevalence of plasmid mediated quinolone resistance determinants qnr and aac(6’)-Ib-cr in Escherichia coli and Klebsiella pneumoniae producing extended-spectrum ß-lactamases in Spain. Int J Antimicrob Agents. 2012;39(5):431–434.
- Seiffert SN, Marschall J, Perreten V, Carattoli A, Furrer H, Endimiani A. Emergence of Klebsiella pneumoniae co-producing NDM-1, OXA 48, CTX-M-15, CMY-16, QnrA and ArmA in Switzerland. Int J Antimicrob Agents. 2014;44(3):260–262.
- Paltansing S, Kraakman MEM, Ras JMC, Wessels E, Bernards AT. Characterization of fluoroquinolone and cephalosporin resistance mechanisms in Enterobacteriaceae isolated in a Dutch teaching hospital reveals the presence of an Escherichia coli ST131 clone with a specific mutation in parE. J Antimicrob Chemother. 2013;68(1):40–45.
- Rodrigues C, Machado E, Ramos H, Peixe L, Novais Â. Expansion of ESBL-producing Klebsiella pneumoniae in hospitalized patients: A successful story of international clones (ST15, ST147, ST336) and epidemic plasmids (IncR, IncFIIK). Int J Med Microbiol. 2014;304(8):1100–1108.
- Beaber JW, Hochhut B, Waldor MK. SOS response promotes horizontal dissemination of antibiotic resistance genes. Nature. 2004;427(6969):72–74.