This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Randegger, C. C.
Right arrow Articles by Hächler, H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Randegger, C. C.
Right arrow Articles by Hächler, H.

 Previous Article  |  Next Article 

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.


This article has been cited by other articles:

  • Andersson, P., Harris, T., Tong, S. Y. C., Giffard, P. M. (2009). Analysis of blaSHV Codon 238 and 240 Allele Mixtures Using Sybr Green High-Resolution Melting Analysis. Antimicrob. Agents Chemother. 53: 2679-2683 [Abstract] [Full Text]  
  • Abou El Hassan, M., Bremner, R. (2009). A rapid simple approach to quantify chromosome conformation capture. Nucleic Acids Res 37: e35-e35 [Abstract] [Full Text]  
  • Jones, C. H., Ruzin, A., Tuckman, M., Visalli, M. A., Petersen, P. J., Bradford, P. A. (2009). Pyrosequencing Using the Single-Nucleotide Polymorphism Protocol for Rapid Determination of TEM- and SHV-Type Extended-Spectrum {beta}-Lactamases in Clinical Isolates and Identification of the Novel {beta}-Lactamase Genes blaSHV-48, blaSHV-105, and blaTEM-155. Antimicrob. Agents Chemother. 53: 977-986 [Abstract] [Full Text]  
  • Naas, T., Oxacelay, C., Nordmann, P. (2007). Identification of CTX-M-Type Extended-Spectrum-{beta}-Lactamase Genes Using Real-Time PCR and Pyrosequencing. Antimicrob. Agents Chemother. 51: 223-230 [Abstract] [Full Text]  
  • Sturenburg, E., Storm, N., Sobottka, I., Horstkotte, M. A., Scherpe, S., Aepfelbacher, M., Muller, S. (2006). Detection and Genotyping of SHV {beta}-Lactamase Variants by Mass Spectrometry after Base-Specific Cleavage of In Vitro-Generated RNA Transcripts.. J. Clin. Microbiol. 44: 909-915 [Abstract] [Full Text]  
  • Espy, M. J., Uhl, J. R., Sloan, L. M., Buckwalter, S. P., Jones, M. F., Vetter, E. A., Yao, J. D. C., Wengenack, N. L., Rosenblatt, J. E., Cockerill, F. R. III, Smith, T. F. (2006). Real-Time PCR in Clinical Microbiology: Applications for Routine Laboratory Testing. Clin. Microbiol. Rev. 19: 165-256 [Abstract] [Full Text]  
  • Szabo, D., Melan, M. A., Hujer, A. M., Bonomo, R. A., Hujer, K. M., Bethel, C. R., Kristof, K., Paterson, D. L. (2005). Molecular Analysis of the Simultaneous Production of Two SHV-Type Extended-Spectrum Beta-Lactamases in a Clinical Isolate of Enterobacter cloacae by Using Single-Nucleotide Polymorphism Genotyping. Antimicrob. Agents Chemother. 49: 4716-4720 [Abstract] [Full Text]  
  • Paterson, D. L., Bonomo, R. A. (2005). Extended-Spectrum {beta}-Lactamases: a Clinical Update. Clin. Microbiol. Rev. 18: 657-686 [Abstract] [Full Text]  
  • Chia, J.-H., Chu, C., Su, L.-H., Chiu, C.-H., Kuo, A.-J., Sun, C.-F., Wu, T.-L. (2005). Development of a Multiplex PCR and SHV Melting-Curve Mutation Detection System for Detection of Some SHV and CTX-M {beta}-Lactamases of Escherichia coli, Klebsiella pneumoniae, and Enterobacter cloacae in Taiwan. J. Clin. Microbiol. 43: 4486-4491 [Abstract] [Full Text]  
  • Weldhagen, G. F. (2004). Rapid Detection and Sequence-Specific Differentiation of Extended-Spectrum {beta}-Lactamase GES-2 from Pseudomonas aeruginosa by Use of a Real-Time PCR Assay. Antimicrob. Agents Chemother. 48: 4059-4062 [Abstract] [Full Text]  
  • Weldhagen, G. F. (2004). Sequence-Selective Recognition of Extended-Spectrum {beta}-Lactamase GES-2 by a Competitive, Peptide Nucleic Acid-Based Multiplex PCR Assay. Antimicrob. Agents Chemother. 48: 3402-3406 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»