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Antimicrobial Agents and Chemotherapy, September 2000, p. 2382-2388, Vol. 44, No. 9
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Characterization of the Extended-Spectrum beta -Lactamase Reference Strain, Klebsiella pneumoniae K6 (ATCC 700603), Which Produces the Novel Enzyme SHV-18

J. Kamile Rasheed,1,* Gregory J. Anderson,1 Hesna Yigit,1 Anne Marie Queenan,2 Antonio Doménech-Sánchez,3 Jana M. Swenson,1 James W. Biddle,1 Mary Jane Ferraro,4 George A. Jacoby,5,6 and Fred C. Tenover1

Hospital Infections Program, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 303331; The R. W. Johnson Pharmaceutical Research Institute, Raritan, New Jersey 088692; Área de Microbiología, Departamento de Biología, Universidad de las Islas Baleares, Palma de Mallorca, Spain3; Microbiology Laboratory, Massachusetts General Hospital, Boston, Massachusetts 021144; Edith Nourse Rogers Memorial Veterans Hospital, Bedford, Massachusetts 017305; and Lahey Clinic, Burlington, Massachusetts 018056

Received 24 August 1999/Returned for modification 13 December 1999/Accepted 30 May 2000

Klebsiella pneumoniae K6 (ATCC 700603), a clinical isolate, is resistant to ceftazidime and other oxyimino-beta -lactams. A consistent reduction in the MICs of oxyimino-beta -lactams by at least 3 twofold dilutions in the presence of clavulanic acid confirmed the utility of K. pneumoniae K6 as a quality control strain for extended-spectrum beta -lactamase (ESBL) detection. Isoelectric-focusing analysis of crude lysates of K6 demonstrated a single beta -lactamase with a pI of 7.8 and a substrate profile showing preferential hydrolysis of cefotaxime compared to ceftazidime. PCR analysis of total bacterial DNA from K6 identified the presence of a blaSHV gene. K6 contained two large plasmids with molecular sizes of approximately 160 and 80 kb. Hybridization of plasmid DNA with a blaSHV-specific probe indicated that a blaSHV gene was encoded on the 80-kb plasmid, which was shown to transfer resistance to ceftazidime in conjugal mating experiments with Escherichia coli HB101. DNA sequencing of this blaSHV-related gene revealed that it differs from blaSHV-1 at nine nucleotides, five of which resulted in amino acid substitutions: Ile to Phe at position 8, Arg to Ser at position 43, Gly to Ala at position 238, and Glu to Lys at position 240. In addition to the production of this novel ESBL, designated SHV-18, analysis of the outer membrane proteins of K6 revealed the loss of the OmpK35 and OmpK37 porins.

* Corresponding author. Mailing address: Nosocomial Pathogens Laboratory Branch (G08), Centers for Disease Control and Prevention, 1600 Clifton Rd. N.E., Atlanta, GA 30333. Phone: (404) 639-3247. Fax: (404) 639-1381. E-mail: Jkr1{at}cdc.gov.

Antimicrobial Agents and Chemotherapy, September 2000, p. 2382-2388, Vol. 44, No. 9
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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