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Antimicrobial Agents and Chemotherapy, August 2000, p. 2230-2230, Vol. 44, No. 8
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
LETTERS TO THE EDITOR
Are SHV 
-Lactamases Universal in Klebsiella
pneumoniae?
 |
LETTER |
The status of SHV-1 or related 
-lactamases as "typical" of
Klebsiella pneumoniae prompted debate at the 39th
Interscience Conference of Antimicrobial Agents and Chemotherapy. To
test whether SHV-type enzymes are ubiquitous in this species, as
previously asserted by one of us, (2) we screened 20 isolates identified as K. pneumoniae with API 20E tests
(BioMerieux, Lyons, France). The isolates were obtained in
1997-1998 from intensive care units across Europe (1) and
were chosen as broadly susceptible to -lactam antibiotics
including aztreonam (MIC 
0.25 µg/ml), ceftazidime (MIC 0.25 µg/ml), ceftazidime plus clavulanic acid, 4 µg/ml (MIC 0.25 µg/ml), cefuroxime (MIC 4 µg/ml),
ceftriaxone (MIC 0.12 µg/ml), piperacillin (MIC 8 µg/ml), piperacillin plus tazobactam, 4 µg/ml (MIC 4 µg/ml), cefoxitin (MIC 8 µg/ml), cefotetan (MIC 0.12 µg/ml), and meropenem (MIC 0.12 µg/ml). The MIC ratio
of ceftazidime to ceftazidime plus clavulanic acid equalled unity,
except for one isolate where it was 2, indicating the absence of
extended-spectrum -lactamases. The isolates were from 16 hospitals
in eight European countries.
Screening for blaSHV genes was by PCR, initially
using primers 5'-TCA GCG AAA AAC ACC TTG-3' and 5'-TCC CGC AGA TAA ATC
ACC A-3' (5) to amplify a 475-bp fragment. These primers
correspond to positions 509 to 526 and 962 to 980, respectively, in the
blaSHV-1 sequence (GenBank accession number
AF124984). Amplification of a DNA fragment of the expected size was
achieved with 18 of the 20 isolates. The two isolates that failed to
give amplification products were reidentified with biochemical tests
(4). One was deduced to be K. oxytoca on the
basis of pectate degradation, utilization of
m-hydroxybenzoate, and the ability to grow at 10°C but was
indole negative and so was not recognized as K. oxytoca by
the API 20E system. The second isolate was confirmed as K. pneumoniae. This isolate was retested for
blaSHV-related DNA using primers
5'-ATGCGTTATATTCGCCTGTG-3' and
5'-GTTAGCGTTGCCAGTGCTCG-3', corresponding to positions 199 to 210 and 1041 to 1060, respectively, of
blaSHV-1. These primers amplified an 865-bp
fragment, corresponding to the entire coding region of
blaSHV.
Phenotypic expression of blaSHV was examined by
isoelectric focusing, performed as described by Livermore and Williams
(3). -Lactamases that cofocused with SHV-1 enzyme (pI
7.6) were detected in 16 of the 19 confirmed K. pneumoniae
strains. No -lactamase band was detected in extracts of the three
remaining isolates, indicating that expression was absent or below the
detection limit of the nitrocefin overlay. These three organisms gave
inhibition zones 3 to 7 mm larger to discs containing amoxicillin plus
clavulanate at 20 + 10 µg than to those containing ampicillin at 25 µg, suggesting that some slight -lactamase activity was present
despite the electrofocusing result. Surprisingly, and despite its lack
of piperacillin resistance, one SHV-1-positive isolate also had a -lactamase that cofocused with TEM-1 enzyme.
We conclude that blaSHV-related genes were
present in all 19 confirmed K. pneumoniae isolates and were
expressed in at least 16 of these strains, which were from diverse
European sources and were selected as highly susceptible to
-lactams. These data support the view that SHV-related enzymes
approach ubiquity in K. pneumoniae, at least in Europe.
| |
ACKNOWLEDGMENTS |
We are grateful to colleagues in the Laboratory of Hospital
Infection for the identification of the two isolates by biochemical tests.
We are grateful to AstraZeneca for financial support.
| |
FOOTNOTES |
*
Phone: 44-020-8200-400
Fax: 44-020-8358-3292
E-mail: Dlivermore{at}phls.nhs.uk
| |
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| | | | |
Gioia S. Babini
David M. Livermore*
Antibiotic Resistance Monitoring & Reference Laboratory
Central Public Health Laboratory
London, United Kingdom, NW9 5HT
|
Antimicrobial Agents and Chemotherapy, August 2000, p. 2230-2230, Vol. 44, No. 8
0066-4804/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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