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Song, Sung, Kwon, Park, Cho, Shin, Ko, Kim, Shin, and Koo: Analysis of Acquired Resistance Genes in Stenotrophomonas maltophilia
Original Article

Korean J Leg Med 2010;30(3):295-300.

Published online: 14 January 2010

DOI: https://doi.org/10.3343/kjlm.2010.30.3.295

Analysis of Acquired Resistance Genes in Stenotrophomonas maltophilia

Jeong Hoon Song, M.D.1, Ji Youn Sung, Ph.D.1, Kye Chul Kwon, M.D.1, Jong Woo Park, M.D.1, Hye Hyun Cho, B.S.1, So Yeon Shin, M.D.1, Young Hyun Ko, M.D.1, Ji Myung Kim, M.D.2, Kyeong Seob Shin, M.D.3, Sun Hoe Koo, M.D.1

1Department of Laboratory Medicine, College of Medicine, Chungnam National University, Daejeon, Korea

2Department of Laboratory Medicine, Eulji University Hospital, Daejeon, Korea

3Department of Laboratory Medicine, College of Medicine, Chungbuk National University, Cheongju, Korea

Corresponding author: Sun Hoe Koo, M.D. Department of Laboratory Medicine, College of Medicine, Chungnam National University, 640 Daesa-dong, Jung-gu, Daejeon 301-747, Korea Tel: +82-42-280-7798, Fax: +82-42-257-5365 E-mail: shkoo@cnu.ac.kr

      Revised 8 March 201029 April 2010       Accepted 30 April 2010

Copyright © 2010 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:

Stenotrophomonas maltophilia is a gram-negative bacillus and a nosocomial pathogen in immunocompromised patients. Trimethoprim/sulfamethoxazole (TMP/SMX) is the drug of choice for treating S. maltophilia infection; however, resistance to TMP/SMX is increasing. In this study, we investigated the relationship between the incidence of TMP/SMX resistance and the presence of sul genes and mobile elements.

Methods:

A total of 120 S. maltophilia isolates were collected from 3 university hospitals between April 2007 and April 2009. Antimicrobial susceptibilities were determined using the disk diffusion method. PCR and DNA sequencing were conducted for the detection of sul1, sul2, class 1 integron, and ISCR2 element. Repetitive extragenic palindromic sequence-based PCR (REP-PCR) was carried out to evaluate the genetic relatedness.

Results:

The TMP/SMX-resistant (R) isolates harbored a significantly higher proportion of sul1 gene and class 1 integron than TMP/SMX-susceptible (S) isolates (P<0.001). Seventeen of 28 isolates with sul1 also had a class 1 integron, but none of the isolates without sul1 had a class 1 integron. The identified gene cassettes within class 1 integrons include aacA4, aadA1, aac6'-II, and qac. None of the 120 isolates carried sul2, glmM, or ISCR2 element. REP-PCR did not show any genetic relatedness among the isolates.

Conclusions:

In Korea, the resistance of S. maltophilia isolates to TMP/SMX is due to sul1 within a class 1 integron rather than to sul2. The class 1 integron also harbors multiple antibiotic resistance genes in addition to sul1, and therefore it could mediate multidrug resistance in S. maltophilia. (Korean J Lab Med 2010;30:295-300)

Keywords: Stenotrophomonas maltophilia, Trimethoprim/sulfamethoxazole, sul1 gene, Class 1 integron

REFERENCES

1.Chang LL., Lin HH., Chang CY., Lu PL. Increased incidence of class 1 integrons in trimethoprim/sulfamethoxazole-resistant clinical isolates of Stenotrophomonas maltophilia. J Antimicrob Chemother. 2007. 59:1038–9.
2.Valdezate S., Vindel A., Loza E., Baquero F., Canton R. Antimicrobial susceptibilities of unique Stenotrophomonas maltophilia clinical strains. Antimicrob Agents Chemother. 2001. 45:1581–4.
3.Ahn GY., Yu FN., Jang SJ., Kim DM., Park G., Moon DS, et al. Pseudooutbreak of Stenotrophomonas maltophilia due to contamination of bronchoscope. Korean J Lab Med. 2007. 27:205–9. (안균열, 유봉남, 장숙진, 김동민, 박 건, 문대수 등. 기관지 내시경 오염에 기인한 Stenotrophomonas maltophilia 의 가유행 발생. 대한진단검사의학회지 2007;27:205-9.).
4.Harlowe HD. Acute mastoiditis following pseudomonas maltophilia infection: case report. Laryngoscope. 1972. 82:882–3.
5.Sutter VL. Identification of Pseudomonas species isolated from hospital environment and human sources. Appl Microbiol. 1968. 16:1532–8.
6.Ben-Tovim T., Eylan E., Romano A., Stein R. Gram-negative bacteria isolated from external eye infections. Infection. 1974. 2:162–5.
crossref
7.Gardner P., Griffin WB., Swartz MN., Kunz LJ. Nonfermentative gramnegative bacilli of nosocomial interest. Am J Med. 1970. 48:735–49.
crossref
8.Seol SY., Jang KS., Jeong OG., Cho ER., Kim NH., Yu HS, et al. Antimicrobial resistance and molecular epidemiologic characteristics of Stenotrophomonas maltophilia isolated from clinical specimen. J Korean Soc Microbiol. 2000. 35:239–50. (설성용, 장경수, 정웅기, 조응래, 김능희, 유학선 등. 병원 재료에서 분리한 Stenotrophomonas maltophilia 의항균제내성및분자역학적특성. 대한미생물학회지 2000;35:239-50.).
9.Chang LL., Chen HF., Chang CY., Lee TM., Wu WJ. Contribution of integrons, and SmeABC and SmeDEF efflux pumps to multidrug resistance in clinical isolates of Stenotrophomonas maltophilia. J Antimicrob Chemother. 2004. 53:518–21.
10.Zhang L., Li XZ., Poole K. SmeDEF multidrug efflux pump contributes to intrinsic multidrug resistance in Stenotrophomonas maltophilia. Antimicrob Agents Chemother. 2001. 45:3497–503.
11.Sader HS., Jones RN. Antimicrobial susceptibility of uncommonly isolated non-enteric Gram-negative bacilli. Int J Antimicrob Agents. 2005. 25:95–109.
crossref
12.Perreten V., Boerlin P. A new sulfonamide resistance gene (sul3) in Escherichia coli is widespread in the pig population of Switzerland. Antimicrob Agents Chemother. 2003. 47:1169–72.
13.Toleman MA., Bennett PM., Bennett DM., Jones RN., Walsh TR. Global emergence of trimethoprim/sulfamethoxazole resistance in Stenotrophomonas maltophilia mediated by acquisition of sul genes. Emerg Infect Dis. 2007. 13:559–65.
14.Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing; sixteenth informational supplement. M100-S18 (M2). Wayne, PA: Clinical and Laboratory Standards Institute;2008.
15.Dillon B., Thomas L., Mohmand G., Zelynski A., Iredell J. Multiplex PCR for screening of integrons in bacterial lysates. J Microbiol Methods. 2005. 62:221–32.
crossref
16.Levesque C., Piche L., Larose C., Roy PH. PCR mapping of integrons reveals several novel combinations of resistance genes. Antimicrob Agents Chemother. 1995. 39:185–91.
crossref
17.Lin CW., Chiou CS., Chang YC., Yang TC. Comparison of pulsed-field gel electrophoresis and three rep-PCR methods for evaluating the genetic relatedness of Stenotrophomonas maltophilia isolates. Lett Appl Microbiol. 2008. 47:393–8.
18.Garrison MW., Anderson DE., Campbell DM., Carroll KC., Malone CL., Anderson JD, et al. Stenotrophomonas maltophilia: emergence of multidrug-resistant strains during therapy and in an in vitro pharmacodynamic chamber model. Antimicrob Agents Chemother. 1996. 40:2859–64.
19.Fedler KA., Biedenbach DJ., Jones RN. Assessment of pathogen frequency and resistance patterns among pediatric patient isolates: report from the 2004 SENTRY Antimicrobial Surveillance Program on 3 continents. Diagn Microbiol Infect Dis. 2006. 56:427–36.
crossref
20.Barbolla R., Catalano M., Orman BE., Famiglietti A., Vay C., Smayevsky J, et al. Class 1 integrons increase trimethoprim-sulfamethoxazole MICs against epidemiologically unrelated Stenotrophomonas maltophilia isolates. Antimicrob Agents Chemother. 2004. 48:666–9.

Fig. 1.
Repetitive extragenic palindromic-PCR (REP-PCR) fingerprints of S. maltophilia isolates. Lanes M1 are 1-kb DNA size markers.
kjlm-30-295f1.tif
Table 1.
Oligonucleotide primers used for PCR amplification and sequencing
Primer pairs Target Sequence (5′-3′) Reference
Int1 F intI1 CAGTGGACATAAGCCTGTTC [15]
Int1 R   CCCGAGGCATAGACTGTA  
Int2 F intI2 GTAGCAAACGAGTGACGAAATG [15]
Int2 R   CACGGATATGCGACAAAAAGGT  
Int3 F intI3 GCCTCCGGCAGCGACTTTCAG [15]
Int3 R   ACGGATCTGCCAAACCTGACT  
LECR2 ISCR2 CACTGGCTGGCAATGTCTAG [13]
RECR2   CTTTGGACCGCAGTTGACTC  
glmR glmM GAGTCAACTGCGGTCCAAAC [13]
glmF   ACGGTATTCGTGGCAAAGCC  
FloF floR TCGACATCCTGGCTTCACTG [13]
FloR   ATTACAAGCGCGACAGTGGC  
Sul1 F sul1 GGATTTTTCTTGAGCCCCGC [1]
Sul1 R   CATTGCCGATCGCGTGAAGT  
Sul2 F sul2 CCTGTTTCGTCCGACACAGA [1]
Sul2 R   GAAGCGCAGCCGCAATTCAT  

Abbreviations: F, forward; R, reverse.

Table 2.
Incidence of trimethoprim/sulfamethoxazole resistance in S. maltophilia and relation to sul1, sul2, glmM, class 1 integron, and ISCR2 element
  Trimethoprim/sulfamethoxazole P value∗
Resistant Susceptible
N of isolates 19 101  
N of isolates with sul1 13 15 <0.001
N of isolates with sul2 0 0  
N of isolates with class 1 integrons 10 7 <0.001
N of isolates with ISCR2 0 0  
N of isolates with glmM 0 0  

∗ Fisher's exact test.

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