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Park, Kang, Bae, Kim, Kim, Uh, Jeong, and Lee: Prevalence of the Extended-Spectrum β-Lactamase and qnr Genes in Clinical Isolates of Escherichia coli
Original Article

Korean J Leg Med 2009;29(3):218-223.

Published online: 14 January 2009

DOI: https://doi.org/10.3343/kjlm.2009.29.3.218

Prevalence of the Extended-Spectrum β-Lactamase and qnr Genes in Clinical Isolates of Escherichia coli

Yongjung Park, M.D.1, Hyun-Kyung Kang, Ph.D.2, Il Kwon Bae, Ph.D.3, Juwon Kim, M.D.1, Jae-Seok Kim, M.D.4, Young Uh, M.D.5, Seok Hoon Jeong, M.D.1, Kyungwon Lee, M.D.1

1Department of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine, Seoul, Korea

2Department of Dental Hygiene, Donju College, Busan, Korea

3Research Institute for Antimicrobial Resistance, Kosin University College of Medicine, Busan, Korea

4Department of Laboratory Medicine, Hallym University College of Medicine, Seoul, Korea

5Department of Laboratory Medicine, Yonsei University Wonju College of Medicine, Wonju, Korea

Corresponding author: Seok Hoon Jeong, M.D. Department of Laboratory Medicine and Research Institute of Bacterial Resistance, Yonsei University College of Medicine, 250 Seongsan-ro, Seodaemun-gu, Seoul 120-752, Korea Tel: +82-2-2228-2448, Fax: +82-2-313-0908 E-mail: kscpjsh@yuhs.ac

      Revised 29 January 20094 March 2009       Accepted 6 March 2009

Copyright © 2009 Korean Society for Legal Medicine

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background:

This study was performed to investigate the prevalence of qnr genes in clinical isolates of Escherichia coli from Korea that produce extended-spectrum β-lactamases (ESBLs).

Methods:

During the period of May to June 2005, we collected clinical isolates of E. coli that were intermediate or resistant to ceftazidime and/or cefotaxime from 11 Korean hospitals. Antimicrobial susceptibility was determined by the disk diffusion and agar dilution methods. ESBL production was confirmed phenotypically by the double-disk synergy test. ESBL and qnr genes were searched for by PCR amplification, and the PCR products were then subjected to direct sequencing.

Results:

Double-disk synergy tests were positive in 84.3% (118/140) of ceftazidime- and/or cefotaxime-nonsusceptible E. coli isolates. The most prevalent types of ESBL in E. coli isolates were CTX-M-14 (N=41) and CTX-M-15 (N=58). Other ESBLs were also identified, including CTX-M-3 (N=7), CTX-M-9 (N=8), CTX-M-12 (N=1), CTX-M-57 (N=1), SHV-2a (N=2), SHV-12 (N=17) and TEM-52 (N= 4). The qnrA1 and qnrB4 genes were identified in 4 and 7 ESBL-producing isolates, respectively.

Conclusions:

CTX-M-type enzymes were the most common type of ESBL in E. coli isolates from Korea, and the qnr genes were not uncommon in ESBL-producing E. coli isolates. Dissemination of E. coli containing both ESBL and qnr genes could compromise the future usefulness of the expanded-spectrum antibiotics for the treatment of infections.

Keywords: Escherichia coli, CTX-M ESBL, qnrA1, qnrB4

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Table 1.
Sequences of the PCR primers
Name Nucleotide Sequence Product size (bp) GenBank Accession No.
TEM F 5′-ctt gaa gac gaa agg gcc tc-3′ 997 M36543
TEM R 5′-tga ctc ccc gtc gtg tag at-3′    
SHV F 5′-cgc cgg att ctt att tg-3′ 1,071 X98100
SHV R 5′-cca cgt tta tgg cgt tac ct-3′    
CTX-M-1F 5′-gga cgt aca gca aaa act tgc-3′ 891 X92506
CTX-M-1R 5′-cgg ttc gct ttc act ttt ctt-3′    
CTX-M-2F 5′-cgg tgc tta aac aga gcg ag-3′ 624 X92507
CTX-M-2R 5′-cca tga ata agc agc tga ttg ccc-3′    
CTX-M-8F 5′-acg ctc aac acc gcg atc-3′ 490 AF189721
CTX-M-8R 5′-cgt ggg ttc tcg ggg ata a-3′    
CTX-M-9F 5′-gat tga ccg tat tgg gag ttt-3′ 947 AJ416345
CTX-M-9R 5′-cgg ctg ggt aaa ata ggt ca-3′    
PER-1 F 5′-gtt aat ttg ggc tta ggg cag-3′ 855 Z21957
PER-1 R 5′-cag cgc aat ccc cac tgt-3′    
VEB F 5′-acc aga tag gag tac aga cat atg a-3 727 AF220758
VEB R 5′-ttc atc acc gcg ata aag cac-3′    
GES F 5′-gtt aga cgg gcg tac aaa gat aat-3′ 903 AY260546
GES R 5′-tgt ccg tgc tca gga tga gt-3′    
TLA F 5′-cgc gaa aat tct gaa atg ac-3′ 992 AF148067
TLA R 5′-agg aaa ttg tac cga gac cct-3′    
qnrA-F 5′-tca gca aga gga ttt ctc acg −3′ 606 DQ989302
qnrA-R 5′-ggt tcc agc agt tgc tcc t-3′    
qnrB1-F 5′-acc tga gcg gca ctg aat tta t-3′ 424 DQ351241
qnrB1-R 5′-tcg caa tgt gtg aag ttt gc-3′    
qnrB4-F 5′-gat gac tct ggc gtt agt tgg-3′ 641 DQ303921
qnrB4-R 5′-cca tga cag cga tac caa ga-3′    
qnrS-F 5′-gac gtg cta act tgc gtg at-3′ 380 DQ449578
qnrS-R 5′-act taa gtc tga ctc ttt cag tga tgc-3′    
Table 2.
Characteristics of Ambler class A ESBL and qnr-producing E. coli isolates
Type of Ambler class A ESBLs (N) Type of qnr genes (N) MIC range (MIC50) (μg/mL)
FOX CAZ CAZ-CLA CTX CTX-CLA CIP
TEM-52 (4)   2-4 16-32 0.25-1 16-32 0.06-0.25 4->256
SHV-2a+CTX-M-14+CTX-M-15 (1) qnrA1 (1) 8 >256 32 >256 128 >256
SHV-2a+CTX-M-15 (1)   4 64 1 256 1 >256
SHV-12 (8) qnrB4 (1) 2-128(8) 32-128 (64) 0.5-32 (2) 2-16 (8) 0.06-8 (0.12) 4->256 (>256)
SHV-12+CTX-M-9 (1) qnrB4 (1) 32 16 0.5 8 0.25 >256
SHV-12+CTX-M-14 (3) qnrA1 (1) 4-256 16-128 0.5-128 16-64 4-128 64->256
SHV-12+CTX-M-15 (5) qnrA1 (1) 2-128 64->256 0.5-32 256->256 0.25-32 >256
CTX-M-3 (6) qnrB4 (1) 2-64 4-32 0.5-32 128-256 1-128 >256
CTX-M-9 (2) qnrB4 (2) 64->256 16-64 8-64 32->256 32-256 4->256
CTX-M-12 (1)   4 1 0.5 32 2 4
CTX-M-14 (31) qnrA1 (1) 1->256 (16) 0.5-64 (4) 0.12-64 (0.5) 4-256 (32) 0.25-128 (2) 2->256 (>256)
  qnrB4 (1)            
CTX-M-14+CTX-M-15 (1)   16 128 4 >256 8 >256
CTX-M-15 (47)   2-32 (8) 16-256 (64) 0.25-128 (2) 128->256 (>256) 0.25-256 (2) 4->256 (>256)
CTX-M-57 (1)   8 32 0.5 256 1 >256

Abbreviations: FOX, cefoxitin; CAZ, ceftazidime; CLA, clavulanic acid; CTX, cefotaxime; CIP, ciprofloxacin.

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