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Sung, Koo, Kwon, Park, Ko, Shin, and Song: Characterization of Class 1 Integrons in Metallo-β-lactamase-producing Pseudomonas aeruginosa
Original Article

Korean J Clin Microbiol 2009;12(1):17-23.

Published online: 22 March 2009

DOI: https://doi.org/10.5145/KJCM.2009.12.1.17

Characterization of Class 1 Integrons in Metallo-β-lactamase-producing Pseudomonas aeruginosa

Ji Youn Sung, Sun Hoe Koo, Kye Chul Kwon, Jong Woo Park, Chi Seon Ko, So Youn Shin, Jeong Hoon Song

Department of Laboratory Medicine, College of Medicine, Chungnam National University, Daejeon, Korea

Correspondence: Sun Hoe Koo, Department of Laboratory Medicine, College of Medicine, Chungnam National University, 640, Daesa-dong, Jung-gu, Daejeon 301-721, Korea. (Tel) 82-42-280-7798, (Fax) 82-42-257-5365, (E-mail) shkoo@cnu.ac.kr

Received 3 July 200813 August 2008       Accepted 20 October 2008

Copyright © 2009 Korean Journal of Clinical Microbiology

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

The genes of metallo-β-lactamase (MBL), a powerful carbapenemase, are carried as a part of the mobile gene cassettes inserted into integrons playing an important role in rapid dissemination of antibiotic resistance genes among bacterial isolates. In this study, we investigated carbapenemase genes and class 1 integrons integrated into the gene cassettes in imipenem-non susceptible P. aeruginosa.

Methods

From July 2006 to March 2008, 81 consecutive, non-duplicate, imipenem-non susceptible P. aeruginosa were isolated at Chungnam National University Hospital in Chungcheong province of Korea. The modified Hodge and double disk synergy tests were conducted for the screening of carbapenemase and MBL production, respectively, and PCR and DNA sequencing were performed for the detection of carbapenemase genes and class 1 integron gene cassettes. We also employed the repetitive element sequence-based (Rep)-PCR method for an epidemiologic study.

Results

MBLs were detected in 13.6% (11/81) of imipenem-non susceptible P. aeruginosa. Ten isolates were found to carry blaIMP-1, whereas 1 isolate was found to carry a blaVIM-2. All of the IMP-1-producing strains harbored 4.0 kb class 1 integron containing chloramphenicol, aminoglycoside, and β-lactam-resistant genes. However, blaIMP-1 was not detected at class 1 integron. A 2.5 kb class 1 integron harboring blaVIM-2 was detected in a VIIM-2- producing strain. One identical pattern was observed in ten IMP-1 producing strains.

Conclusion

IMP-1 producing P. aeruginosa strains are currently distributed throughout Chungcheong province of Korea. In particular, all of the strains harbored class 1 integrons containing variant antibiotic resistance gene cassettes.

Keywords: Metallo-β-lactamase (MBL), Class 1 integron, Multidrug resistance Pseudomonas aeruginosa

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Fig. 1.
Schematic representation of gene cassette structure located in the class 1 integron isolated from Pseudomonas aeruginosa. The horizontal arrows indicate the translation orientation of the genes.
kjcm-12-17f1.tif
Fig. 2.
Repetitive element sequence-based (Rep)-PCR patterns of genomic DNA from eleven MBL producing Pseudomonas aeruginosa. Lane M is 1 kb DNA size marker.
kjcm-12-17f2.tif
Table 1.
Oligonucleotide primers for amplification and sequencing
Enzyme class Primer pairs Target Sequence (5’-3’) Amplicon size (bp) Reference
Class A PER1 F blaPER1 GTTAATTTGGGCTTAGGGCAGA 855 22
PER1 R CAGCGCAATCCCCACTGT
PSE F blaPSE AATGGCAATCAGCGCTTC 700 23
PSE R GCGCGACTGTGATGTATA
VEB F blaVEB CGACTTCCATTTCCCGATGC 650 24
VEB R GGACTCTGCAACAAATACGC
GES F blaGES, blaIBC GTTAGACGGGCGTACAAAGATAAT 903 25
GES R TGTCCGTGCTCAGGATGAGT
Class B IMP F blaIMP CATGGTTTGGTGGTTCTTGT 488 26
IMP R ATAATTTGGCGGACTTTGGC
VIM F blaVIM ATTGGTCTATTTGACCGCGTC 780 26
VIM R TGCTACTCAACGACTGAGCG
SIM F blaSIM GTACAAGGGATTCGGCATCG 569 27
SIM R TGGCCTGTTCCCATGTGAG
SPM F blaSPM CTAAATCGAGAGCCCTGCTTG 798 27
SPM R CCTTTTCCGCGACCTTGATC
GIM F blaGIM TCAATTAGCTCTTGGGCTGAC 72 27
GIM R CGGAACGACCATTTGAATGG
Class D OXA-23F blaOXA-23, 27, 49 GATGTGTCATAGTATTCGTCG 1,058 26
OXA-23R TCACAACAACTAAAAGCACTG
OXA-24F blaOXA-24, 25, 26, 40, 72 GTACTAATCAAA GTTGTGAA 825 26
OXA-24R TTCCCCTAACATGAATTTGT
OXA-51F blaOXA-51like TGAACATTAAAICACTCTT 825 28
OXA-51R CTATAAAATACCTAATTGTT
OXA-58F blaOXA-58 CGATCAGAATGTTCAAGCGC 528 29
OXA-58R ACGATTCTCCCCTCTGCGC

Abbreviations: F, forward; R, reverse.

Table 2.
Antimicrobial susceptibilities of Pseudomonas aeruginosa isolates
Antimicrobial agents % susceptibility
Total isolated P. aeruginosa (n=81) IMP-1 producing P. aeruginosa (n=10)
Intermediate Resistant Intermediate Resistant
Amikacin
Gentamicin
Netilmicin
Tobramycin
Aztreonam
Ceftazidime
Cefepime
Imipenem
Meropenem
Piperacillin
Ticarcillin
Ciprofloxacin		 3.7
14.8
12.3
0.0
28.4
12.3
19.8
23.5
30.9
0.0
0.0
2.5		 56.8
70.4
65.4
60.5
42.0
54.3
51.9
76.5
48.1
43.2
69.1
76.5		 0.0
0.0
0.0
0.0
70.0
0.0
0.0
0.0
0.0
0.0
0.0
0.0		 100.0
100.0
100.0
100.0
30.0
100.0
100.0
100.0
100.0
60.0
100.0
100.0
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