Antimicrobial Agents and Chemotherapy, June 2001, p. 1730-1736, Vol. 45, No. 6
0066-4804/01/$04.00+0 DOI: 10.1128/AAC.45.6.1730-1736.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
-Lactamases
Institute of Medical Microbiology, University of Zürich, Zürich, Switzerland
Received 30 October 2000/Returned for modification 26 February 2001/Accepted 16 March 2001
Extended-spectrum
-lactamases (ESBLs), e.g., ESBLs of the TEM or
SHV type, compromise the efficacies of expanded-spectrum cephalosporins. An SHV non-ESBL that hydrolyzes only narrow-spectrum cephalosporins can be converted into an SHV ESBL through substitutions at three amino acid positions, 179, 238, or 238-240. In order to
improve detection of SHV ESBLs, a novel method, based on real-time PCR
monitored with fluorescently labeled hybridization probes and followed
by melting curve analysis, was developed. It is able to (i) detect
blaSHV genes with high degrees of sensitivity
and specificity, (ii) discriminate between blaSHV
non-ESBL and blaSHV ESBL, and (iii)
categorize the SHV ESBL producers into three phenotypically relevant
subgroups. This method, termed the SHV melting curve mutation detection
method, represents a powerful tool for epidemiological studies with SHV
ESBLs. It even has the potential to be used in the diagnostic
microbiology laboratory, because up to 32 clinical isolates can be
processed in less than 1 h by starting with just a few bacterial colonies.
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