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Nosocomial spread of multi-resistant Klebsiella pneumoniae containing a plasmid encoding multiple ß-lactamases

Infectious Disease Dept, The First Affiliated Hospital, Medical School, Zhejiang University, The Key Laboratory of Infectious Diseases of Public Health Ministry, Zhejiang, Hangzhou, China

Correspondence Yun-Song Yu yvys119{at}163.com

Received May 10, 2005
Accepted June 17, 2005

Six Klebsiella pneumoniae isolates that exhibited resistance to a wide spectrum of antibiotics were recovered from the intensive care units in the First Affiliated Hospital, Zhejiang University, Hangzhou, China. All isolates contained two plasmids of approximately 95 kb and 200 kb. The 95 kb plasmid was shown to be transferable by conjugation experiments. Isoelectric focusing patterns of the ß-lactamases extracted from the six transconjugants were identical, displaying five pI bands: 5.4, 7.75, 8.0, 8.2 and 8.4. The band corresponding to a pI of 7.75 could be inhibited by cloxacillin but not clavulanic acid, while the other bands could be inhibited by clavulanic acid but not cloxacillin. The 95 kb plasmid was digested with HindIII and a recombinant plasmid pT948 was obtained. The insert was found to contain bla_DHA–1, regulatory gene ampR and an insertion element (IS26), which was downstream of bla_DHA–1. PCR and DNA sequencing results confirmed that the 95 kb plasmid encoded at least four ß-lactamase genes: bla_TEM–1, bla_SHV–12, bla_CTX–M–3 and bla_DHA–1. Epidemiological typing by PFGE of the six clinical isolates of K. pneumoniae demonstrated identical genotypic patterns. In conclusion, all results indicated that the six multi-drug resistant clinical isolates of K. pneumoniae most probably originated from one clone and caused a localized epidemic in the intensive care units.

The GenBank/EMBL/DDBJ accession number for the DHA-1 sequence described in this study is AY705809.

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