Skip to main page content

• HOME
• Current Issue
• Archive
• Alerts
• About ASM
• Contact us
• Tech Support
• Journals.ASM.org

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Jeong, S. H. Articles by Lee, S. H. Search for Related Content PubMed PubMed Citation Articles by Jeong, S. H. Articles by Lee, S. H.

 Previous Article  |  Next Article 

Antimicrobial Agents and Chemotherapy, November 2005, p. 4809-4810, Vol. 49, No. 11
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.11.4809-4810.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

LETTERS TO THE EDITOR

First Outbreak of Klebsiella pneumoniae Clinical Isolates Producing GES-5 and SHV-12 Extended-Spectrum ß-Lactamases in Korea

Top Letter References

 
Klebsiella pneumoniae clinical isolates producing extended-spectrum ß-lactamases (ESBLs; TEM and SHV types) are frequently implicated in hospital outbreaks. There are also reports of K. pneumoniae isolates producing various non-TEM, non-SHV ESBLs: CTX-M and GES/IBC types (1). Presently, the different GES-type ESBLs are designated by identical names (8). To clarify the misleading nomenclature of GES-type ESBLs, we propose maintaining the current denomination concerning the fully characterized GES-3 and GES-4 ESBLs reported by Wachino et al. (12, 13) and renaming the variants (GES-3 and GES-4) reported by Vourli et al. (11) as GES-5 and GES-6, respectively.

In 2004, six K. pneumoniae clinical isolates producing ESBLs were collected from different patients and distributed among several wards (intensive care unit, neurosurgery, pulmonary internal medicine, and general medicine) at Bundang CHA Medical Center, Republic of Korea. These isolates were identified from sputum samples (for three isolates), urine (for one isolate), bile (for one isolate), and pus (for one isolate).

Antibiotic susceptibility testing by disk diffusion tests that were performed according to the recommendations of the Clinical and Laboratory Standards Institute (2) with BBL (Cockeysville, Md.) disks and by the double-disk synergy test of Jarlier et al. (4) suggested the presence of ESBL(s). Double-disk synergies were observed for amoxicillin-clavulanic acid (CLA), ceftazidime, cefotaxime, and aztreonam. All isolates were resistant to penicillins, ceftazidime, cefotaxime, aztreonam, tobramycin, gentamicin, and trimethoprim-sulfamethoxazole. They were susceptible to cefepime and represented reduced susceptibility to imipenem.

The analysis of genomic DNA, digested with SpeI and resolved by pulsed-field gel electrophoresis (PFGE) (5), revealed the same macrorestriction pattern among all isolates, which were therefore classified as indistinguishable.

Screening for ESBL gene(s) was performed by PCR using the primers for bla_TEM, bla_SHV, bla_GES, bla_PER, bla_VEB, bla_TOHO, bla_SFO, bla_BES, bla_FEC, bla_CME, bla_TLA, bla_CTX-M, and bla_OXA (1, 5, 7, 10). PCR amplification experiments using primers designed to amplify GES- and SHV-type ESBL genes gave only positive results for all isolates. The nucleotide sequence of the respective amplicon for the bla_SHV gene revealed 100% identity with bla_SHV-12 from K. pneumoniae (6). That for the bla_GES gene differs by only one silent mutation (GA) at position 54 from bla_GES-5, described elsewhere for Escherichia coli (11). On the isoelectric focusing gel, two ß-lactamase activities with pIs of 5.8 and 8.2 were detected in all isolates. On the basis of DNA sequencing and pI values, the ß-lactamase activity of pI 8.2 corresponds to that of SHV-12 ß-lactamase, and the pI value of 5.8 represents GES-5 ß-lactamase.

Six isolates were used as donors in transconjugation experiments. Ceftazidime-resistant transconjugants were obtained in all cases at frequencies ranging from 10^–4 to 10^–5 per donor cell. All transconjugants produced only GES-5. The SHV-12 gene was not cotransferred, indicating that the bla genes for GES-5 and SHV-12 resided on different genetic elements. All GES-5-producing transconjugants were resistant to penicillins and ceftazidime and susceptible to cefoxitin, cefotaxime, and aztreonam. This phenotype is characteristic of GES/IBC-producing Enterobacteriaceae (9-13). All transconjugants were susceptible to cefepime and showed reduced susceptibilities to imipenem. MICs of ticarcillin, ceftazidime, and cefotaxime were reduced by clavulanic acid (Table 1). This resistance was due to the acquisition of large (>70-kb) plasmids that exhibited identical HindIII restriction patterns and harbored the GES-5 ESBL gene. Due to the presence of SHV-12 in all isolates, the MIC patterns of clinical isolates were different from those of their transconjugants.

View this table: [in this window] [in a new window]

TABLE 1. MICs of ß-lactams for ESBL-producing K. pneumoniae clinical isolates, their transconjugants, and the azide-resistant E. coli J53 recipient

The carbapenemase bioassay (the modified cloverleaf test) (5) showed that extracts derived from all transconjugants producing GES-5 supported growth of the indicator, E. coli ATCC 25922, in the presence of an imipenem disk (30 µg), resembling the effect of OXA-23, which was used as a positive control. In contrast, cell extracts of a ceftazidime-sensitive K. pneumoniae clinical isolate collected from the Bundang CHA Medical Center did not cause any visible inhibition of carbapenemase activity. These findings indicated that the reduced susceptibilities of all isolates and their transconjugants to imipenem were due to production of GES-5 ESBL with the detectable carbapenemase activity.

Finding the same macrorestriction pattern by PFGE indicated that an endemic K. pneumoniae strain producing GES-5 and SHV-12 ESBLs was present in different wards at the Bundang CHA Medical Center in 2004. Six GES-5-producing transconjugants were also resistant to tobramycin, gentamicin, and trimethoprim-sulfamethoxazole. Although the complete nucleotide sequences of integrons from six clinical isolates were not determined, screening for the presence of class 1 through 4 integrase genes from large plasmids by PCR with primer pairs previously described (3) revealed the presence of class 1 integrons. The complete class 1 integron mapping of large plasmids from six clinical isolates and the possible spread of the GES-5 ESBL gene to other Korean hospitals are currently under investigation.

 
This work was funded by a Korea Research Foundation grant (KRF-2004-042-E00117).

Top Letter References

 

1
1. Bradford, P. A. 2001. Extended-spectrum ß-lactamases in the 21st century: characterization, epidemiology and detection of this important resistance threat. Clin. Microbiol. Rev. 14:933-951.[Abstract/Free Full Text]
2
2. Clinical and Laboratory Standards Institute. 2005. Performance standards for antimicrobial disk susceptibility tests, 8th ed. Approved standard M2-A8. Clinical and Laboratory Standards Institute, Wayne, Pa.
3
3. Goldstein, C., M. D. Lee, S. Sanchez, C. Hudson, B. Phillips, B. Register, M. Grady, C. Liebert, A. O. Summers, D. G. White, and J. J. Maurer. 2001. Incidence of class 1 and 2 integrases in clinical and commensal bacteria from livestock, companion animals, and exotics. Antimicrob. Agents Chemother. 44:723-726.
4
4. Jarlier, V., M.-H. Nicolas, G. Fournier, and A. Philippon. 1988. Extended broad-spectrum ß-lactamases conferring transferable resistance to newer ß-lactam agents in Enterobacteriaceae: hospital prevalence and susceptibility patterns. Rev. Infect. Dis. 10:867-878.[Medline]
5
5. Jeon, B.-C., S. H. Jeong, I. K. Bae, S. B. Kwon, K. Lee, D. Young, J. H. Lee, J. S. Song, and S. H. Lee. 2005. Investigation of a nosocomial outbreak of imipenem-resistant Acinetobacter baumannii producing the OXA-23 ß-lactamase in Korea. J. Clin. Microbiol. 43:2241-2245.[Abstract/Free Full Text]
6
6. Jeong, S. H., I. K. Bae, J. H. Lee, S. H. Sohn, G. H. Kang, G. J. Jeon, Y. H. Kim, B. C. Jeong, and S. H. Lee. 2004. Molecular characterization of extended-spectrum ß-lactamases produced by clinical isolates of Klebsiella pneumoniae and Escherichia coli from a Korean nationwide survey. J. Clin. Microbiol. 42:2902-2906.[Abstract/Free Full Text]
7
7. Lee, S. H., J. Y. Kim, S. H. Shin, S. K. Lee, M. M. Choi, I. Y. Lee, Y. B. Kim, J. Y. Cho, W. Jin, and K. J. Lee. 2001. Restriction fragment length dimorphism-PCR method for the detection of extended-spectrum ß-lactamases unrelated to TEM- and SHV-types. FEMS Microbiol. Lett. 200:157-161.[Medline]
8
8. Lee, S. H., S. H. Jeong, J.-I. Wachino, Y. Arakawa, L. Poirel, and P. Nordmann. 2005. Nomenclature of GES-type extended-spectrum ß-lactamases. Antimicrob. Agents Chemother. 49:2148-2150.[Free Full Text]
9
9. Poirel, L., G. F. Weldhagen, T. Naas, C. D. Champs, M. G. Dove, and P. Nordmann. 2001. GES-2, a class A ß-lactamase from Pseudomonas aeruginosa with increased hydrolysis of imipenem. Antimicrob. Agents Chemother. 45:2598-2603.[Abstract/Free Full Text]
10
10. Poirel, L., I. L. Thomas, T. Naas, A. Karim, and P. Nordmann. 2000. Biochemical sequence analyses of GES-1, a novel class A extended-spectrum ß-lactamase, and the class 1 integron IN52 from Klebsiella pneumoniae. Antimicrob. Agents Chemother. 44:622-632.[Abstract/Free Full Text]
11
11. Vourli, S., P. Giakkoupi, V. Miriagou, E. Tzelepi, A. C. Vatopoulos, and L. S. Tzouvelekis. 2004. Novel GES/IBC extended-spectrum ß-lactamase variants with carbapenemase activity in clinical enterobacteria. FEMS Microbiol. Lett. 234:209-213.[Medline]
12
12. Wachino, J.-I., Y. Doi, K. Yamane, N. Shibata, T. Yagi, T. Kubota, H. Ito, and Y. Arakawa. 2004. Nosocomial spread of ceftazidime-resistant Klebsiella pneumoniae strains producing a novel class A ß-lactamase, GES-3, in a neonatal intensive care unit in Japan. Antimicrob. Agents Chemother. 48:1960-1967.[Abstract/Free Full Text]
13
13. Wachino, J.-I., Y. Doi, K. Yamane, N. Shibata, T. Yagi, T. Kubota, and Y. Arakawa. 2004. Molecular characterization of a cephamycin-hydrolyzing and inhibitor-resistant class A ß-lactamase, GES-4, possessing a single G170S substitution in the -loop. Antimicrob. Agents Chemother. 48:2905-2910.[Abstract/Free Full Text]

						Seok Hoon Jeong Il Kwon Bae Doelman Kim Department of Laboratory Medicine College of Medicine Kosin University Busan 602-702 Republic of Korea Seong Geun Hong Department of Laboratory Medicine Pochon CHA University Pochon Republic of Korea Jae Seok Song Jung Hun Lee Sang Hee Lee* Department of Biological Sciences Myongji University San 38-2 Namdong Yongin Kyunggido 449-728 Republic of Korea
						* Phone: 82 31 330 6195, Fax: 82 31 335 8249, E-mail: sangheelee{at}mju.ac.kr

---

Antimicrobial Agents and Chemotherapy, November 2005, p. 4809-4810, Vol. 49, No. 11
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.11.4809-4810.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

This article has been cited by other articles:

• Poirel, L., Carrer, A., Pitout, J. D., Nordmann, P. (2009). Integron Mobilization Unit as a Source of Mobility of Antibiotic Resistance Genes. Antimicrob. Agents Chemother. 53: 2492-2498 [Abstract] [Full Text]  
• Carrer, A., Poirel, L., Eraksoy, H., Cagatay, A. A., Badur, S., Nordmann, P. (2008). Spread of OXA-48-Positive Carbapenem-Resistant Klebsiella pneumoniae Isolates in Istanbul, Turkey. Antimicrob. Agents Chemother. 52: 2950-2954 [Abstract] [Full Text]  
• Walther-Rasmussen, J., Hoiby, N. (2007). Class A carbapenemases. J Antimicrob Chemother 60: 470-482 [Abstract] [Full Text]  
• Queenan, A. M., Bush, K. (2007). Carbapenemases: the Versatile {beta}-Lactamases. Clin. Microbiol. Rev. 20: 440-458 [Abstract] [Full Text]  
• da Fonseca, E. L., Vieira, V. V., Cipriano, R., Vicente, A. C. P. (2007). Emergence of blaGES-5 in clinical colistin-only-sensitive (COS) Pseudomonas aeruginosa strain in Brazil. J Antimicrob Chemother 59: 576-577 [Full Text]  
• Poirel, L., Naas, T., Nordmann, P. (2006). Pyrosequencing as a Rapid Tool for Identification of GES-Type Extended-Spectrum {beta}-Lactamases.. J. Clin. Microbiol. 44: 3008-3011 [Abstract] [Full Text]  

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Jeong, S. H. Articles by Lee, S. H. Search for Related Content PubMed PubMed Citation Articles by Jeong, S. H. Articles by Lee, S. H.