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Antimicrobial Agents and Chemotherapy, June 2008, p. 2014-2018, Vol. 52, No. 6
0066-4804/08/$08.00+0     doi:10.1128/AAC.01539-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Emergence of Serratia marcescens, Klebsiella pneumoniae, and Escherichia coli Isolates Possessing the Plasmid-Mediated Carbapenem-Hydrolyzing β-Lactamase KPC-2 in Intensive Care Units of a Chinese Hospital{triangledown}

Jia Chang Cai, Hong Wei Zhou, Rong Zhang, and Gong-Xiang Chen*

2nd Affiliated Hospital of Zhejiang University, Zhejiang University, 88 Jiefang Rd., Hangzhou, China 310009

Received 28 November 2007/ Returned for modification 25 January 2008/ Accepted 5 March 2008

Twenty-one Serratia marcescens, ten Klebsiella pneumoniae, and one Escherichia coli isolate with carbapenem resistance or reduced carbapenem susceptibility were recovered from intensive care units (ICUs) in our hospital. Enterobacterial repetitive intergenic consensus-PCR and pulsed-field gel electrophoresis demonstrated that all the S. marcescens isolates belonged to a clonal strain and the 10 K. pneumoniae isolates were indistinguishable or closely related to each other. The MICs of imipenem, meropenem, and ertapenem for all isolates were 2 to 8 µg/ml, except for K. pneumoniae K10 (MICs of 128, 256, and >256 µg/ml). Isoelectric focusing, PCRs, and DNA sequencing indicated that all S. marcescens isolates produced KPC-2 and a β-lactamase with a pI of 6.5. All K. pneumoniae isolates produced TEM-1, KPC-2, CTX-M-14, and a β-lactamase with a pI of 7.3. The E. coli E1 isolate produced KPC-2, CTX-M-15, and a β-lactamase with a pI of 7.3. Conjugation studies with E. coli (EC600) resulted in the transfer of reduced carbapenem susceptibility compared to that of the original isolates, and only the blaKPC-2 gene was detected in E. coli transconjugants. Plasmid restriction analysis showed identical restriction patterns among all E. coli transconjugants. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and ompK35/36 gene sequence analysis of outer membrane proteins revealed that K. pneumoniae K10 failed to express OmpK36, because of insertional inactivation by an insertion sequence ISEcp1. All these results indicate that KPC-2-producing S. marcescens, K. pneumoniae, and E. coli isolates emerged in ICUs in our hospital. KPC-2 combined with porin deficiency results in high-level carbapenem resistance in K. pneumoniae. The same blaKPC-2-encoding plasmid was spread among the three different genera.

* Corresponding author. Mailing address: 2nd Affiliated Hospital of Zhejiang University, 88 Jiefang Rd., Hangzhou, China 310009. Phone: 86-571-8778-4633. Fax: 86-571-8673-6937. E-mail: chengong218{at}163.com

{triangledown} Published ahead of print on 10 March 2008.

Antimicrobial Agents and Chemotherapy, June 2008, p. 2014-2018, Vol. 52, No. 6
0066-4804/08/$08.00+0     doi:10.1128/AAC.01539-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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