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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.

Real-Time PCR and Melting Curve Analysis for Reliable and Rapid Detection of SHV Extended-Spectrum beta -Lactamases

Corinne C. Randegger and Herbert Hächler*

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 beta -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.

* Corresponding author. Mailing address: Institute of Medical Microbiology, University of Zürich, P.O. Box, CH-8028 Zürich, Switzerland. Phone: 41-1-634-2648. Fax: 41-1-634-4906. E-mail: haechler{at}immv.unizh.ch.

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.


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