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Sung, Kwon, Park, Kim, Kim, Shin, Kim, Ko, Shin, Song, and Koo: Dissemination of IMP-1 and OXA Type β-Lactamase in Carbapenem-resistant Acinetobacter baumannii
Original Article

Korean J Leg Med 2008;28(1):16-23.

Published online: 14 February 2008

DOI: https://doi.org/10.3343/kjlm.2008.28.1.16

Dissemination of IMP-1 and OXA Type β-Lactamase in Carbapenem-resistant Acinetobacter baumannii

Ji Youn Sung1, Kye Chul Kwon, M.D.1, Jong Woo Park, M.D.1, Yeon Suk Kim, M.D.2, Ji Myung Kim, M.D.3, Kyeong Seob Shin, M.D.4, Jong Wan Kim, M.D.5, Chi Seon Ko, M.D.1, So Youn Shin, M.D.1, Jeong Hoon Song, M.D.1, Sun Hoe Koo, M.D.1

1Department of Laboratory Medicine, and Division of Infectious Diseases, Daejeon, Korea

2Department of Internal Medicine, Chungnam National University Hospital, Daejeon, Korea

3Department of Laboratory Medicine, Eulji University School of Medicine, Daejeon, Korea

4Department of Laboratory Medicine, Chungbuk National University College of Medicine, Cheongju, Korea

5Department of Laboratory Medicine, Dankook University College of Medicine, Cheonan, Korea

∗This work was supported by grant no. R11-2002-100-00000-0 from the ERC Program of the Korea Science & Engineering Foundation and in part by the grant from Chungnam university hospital, 2005.

Received 11 September 200713 November 2007       Accepted 13 November 2007

Copyright © 2008 by the 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

Acinetobacter baumannii is an aerobic, gram-negative, glucose-nonfermenting bacterium, which has emerged as a serious opportunistic pathogen. In recent years, the increasing instance of carbapenem-resistant A. baumannii producing metallo-β-lactamases (MBLs) or OXA-type β-lactamases is causing a serious clinical problem. In this study, we investigated the prevalence of Ambler class A, B, and D β-lactamases and their extended-spectrum derivatives in carbapenem-resistant A. baumannii isolates.

Methods

A total of 31 consecutive, non-duplicate, carbapenem-resistant A. baumannii were isolated from three university hospitals in the Chungcheong province of Korea. The modified Hodge and inhibitor-potentiated disk diffusion tests were conducted for the screening of carbapenemase and MBL production, respectively. PCR and DNA sequencing were performed for the detection of β-lactamase genes. We also employed the enterobacterial repetitive intergenic consensus (ERIC)-PCR method for the epidemiologic study.

Results

Twenty-three of 31 isolates harbored blaOXA-2 (51.6%), blaOXA-23 (22.6%), blaIMP-1 (48.4%), and blaVIM-2 (3.2%). All of the OXA-2-producing strains also evidenced MBLs. The strains that harbored blaOXA-23 were isolated only in hospital C, and only in a limited fashion. The ERIC-PCR pattern of the five OXA-23 strains indicated that the isolates were closely related in terms of clonality. The six strains producing IMP-1 isolated from hospital A were confirmed to be identical strains.

Conclusions

A. baumannii strains harboring IMP-1 or OXA-type β-lactamases are currently widely distributed throughout the Chungcheong province of Korea. The most notable finding in this study was that a blaOXA-2-producing A. baumannii harboring MBL, which has not been previously reported, can also lead to outbreaks.

Keywords: A. baumannii, MBL, OXA-2

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Fig. 1.
ERIC-PCR patterns of genomic DNA from seven clinical isolates of A. baumannii harboring blaoxa-23. Lane M1 and M2 are 1 kb and 100 bp DNA size markers, respectively.
kjlm-28-16f1.tif
Fig. 2.
ERIC-PCR patterns of genomic DNA from ten clinical isolates of A. baumannii harboring blaIMP-1. Lane M1 is a 1 kb DNA size marker.
kjlm-28-16f2.tif
Table 1.
Oligonucleotides used as primers for amplification and sequencing in this study
Class Primer pairs Target Sequence (5′→3′) Amplicon size (bp) Reference
Class A TEM F TEM-1 and derivative ATG AGT ATT CAA CAT TTC CGT 861 24
  TEM R   TTA CCA ATG CTT AAT CAG TGA    
  SHV F SHV-1 and derivative CCG GGT TAT TCT TAT TTG TCG CT 831 24
  SHV R   TAG CGT TGC CAG TGC TCG    
  CTX-M F CTX-M-1, 2, 9 group GAT TGA CCG TAT TGG GAG TTT 947 26
  CTX-M R   CGG CTG GGT AAA ATA GGT CA    
  PER F PER-1 GTT AAT TTG GGC TTA GGG CAGA 855 26
  PER R   CAG CGC AAT CCC CAC TGT    
  VEB F VEB-1 CGA CTT CCA TTT CCC GAT GC 650 25
  VEB R   GGA CTC TGC AAC AAA TAC GC    
  GES F GES-1, −2, 3, 4, IBC-1 GTT AGA CGG GCG TAC AAA GAT AAT 903 24
  GES R   TGT CCG TGC TCA GGA TGA GT    
  PSE F PSE-1 AAT GGC AAT CAG CGC TTC 700 22
  PSE R   GCG CGA CTG TGA TGT ATA    
Class B IMP F IMP CAT GGT TTG GTG GTT CTT GT 488 20
  IMP R   ATA ATT TGG CGG ACT TTG GC    
  VIM F VIM ATT GGT CTA TTT GAC CGC GTC 780 20
  VIM R   TGC TAC TCA ACG ACT GAG CG    
Class D OXA-1F OXA group III AGC CGT TAA AAT TAA GCC C 908 11
  OXA-1R   CTT GAT TGA AGG GTT GGG CG    
  OXA-2F OXA group II GCC AAA GGC ACG ATA GTT GT 700 22
  OXA-2R   GCG TCC GAG TTG ACT GCC GG    
  OXA-10F OXA group I TCT TTC GAG TAC GGC ATT AGC 760 25
  OXA-10R   CCA ATG ATG CCC TCA CTT TCC    
  OXA-23F OXA 23, 27, 49 GAT GTG TCA TAG TAT TCG TCG 1,058 20
  OXA-23R   TCA CAA CAA CTA AAA GCA CTG    
  OXA-24F OXA 24, 25, 26, 40, 72 GTA CTA ATC AAA GTT GTG AA 825 20
  OXA-24R   TTC CCC TAA CAT GAA TTT GT    
  OXA-58F OXA 58 CGA TCA GAA TGT TCA AGC GC 528 23
  OXA-58R   ACG ATT CTC CCC TCT GCG C    

Abbreviations: F, forward; R, reverse.

Table 2.
Characterization of carbapenem-resistant A. baumannii isolates
Isolates Antibiotic susceptibilities Clover-leaf IPD β-lactamase
AMK GEN TOB ATM CAZ FEP IPM MEM PIP TZP CIP
A1 R R R R R R R R R I R + + IMP-1, OXA-2
A3 R R R R R R R R R I R + + IMP-1, OXA-2
A4 R R R R R R R R R I R - -  
A5 S S I I R R R R R S S + + IMP-1, OXA-2
A6 S R I R R R R R R S S + + IMP-1, OXA-2
A7 R R R R R R R R R I R + + IMP-1, OXA-2
A8 S R R R R R R R R S S + + IMP-1, OXA-2
A9 R R R R R R R R I S R + + IMP-1, OXA-2
A10 S R R R R R R R R R R + + IMP-1, OXA-2
A11 S S I R R R R R R S S + + IMP-1, OXA-2
A12 S R R R R R R R R S S + + IMP-1, OXA-2
A14 R R R R R R R R R R R - -  
A15 S S S R R R R R R R S - -  
A16 S R R R R R R R R R I - -  
B25 R R R I R R R R R S S + + IMP-1, OXA-2
B75 R R R I R R R R S S I + + IMP-1, OXA-2
B297 R R R I R R R R R S S + + IMP-1, OXA-2
B814 R R R R R S R R S R S + + VIM-2, OXA-2
B852 R R R I R I R R R S I + + IMP-1, OXA-2 OXA-23
C1 R R R R R R R R R R R + -  
C2 R R R R R R R R R I R + + IMP-1, OXA-2
C3 R R R R R R R R R R R - -  
C4 R R R R R R R R R R R + - OXA-23
C5 R R R R R R R R R R R - -  
C6 R R R R R R R R R R R + - OXA-23
C9 R R R R R R R R R R R + - OXA-23
C13 R R R R R R R R R R R + - OXA-23
C14 R R R R R R R R R I R - -  
C15 R R R R R R R R R R R + - OXA-23
C16 R R R R R R R R R R R + - OXA-23
C17 R R R R R R R R R I R - -  
19606∗                       - -  
YMC†                       + + VIM-2

∗ Non-carbapenemase producing A. baumanniiATCC 19606,

† VIM-2-producing A. genomospecies3 YMC 99/11/160.

Abbreviations: AMK, amikacin; GEN, gentamicin; TOB, tobramycin; ATM, aztreonam; CAZ, ceftazidime; FEP, cefepime; IPM, imipenem; MEM, meropenem; PIP, piperacillin; TZP, piperacillin-tazobactam; CIP, ciprofloxacin

Table 3.
Prevalence of β-lactamases in carbapenem- resistant A. baumannii isolates
Class of β-lactamase Type of β-lactamase N (%) of isolates N (%) of isolates in each hospital
A B C
Class A TEM 0 0 0 0
  SHV 0 0 0 0
  CTX-9 0 0 0 0
  PER-1 0 0 0 0
  VEB 0 0 0 0
  GES/IBC 0 0 0 0
  PSE 0 0 0 0
Class B IMP-1 15 (48.4) 10 (32.3) 4 (12.9) 1 (3.2)
  VIM-2 1 (3.2) 0 1 (3.2) 0
Class D OXA-1 0 0 0 0
  OXA-2 16 (51.6) 10 (32.3) 5 (16.1) 1 (3.2)
  OXA-10 0 0 0 0
  OXA-23 7 (22.6) 0 0 7 (22.6)
  OXA-24 0 0 0 0
  OXA-58 0 0 0 0
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