Sequence-selective recognition of extended-spectrum beta-lactamase GES-2 by a competitive, peptide nucleic acid-based multiplex PCR assay.
Extended-spectrum beta-lactamases (ESBLs) in Pseudomonas aeruginosa, such as GES-2, which compromises the efficacy of imipenem, tend to be geographically restricted. The CC-to-AA base pair substitution at positions 493 and 494 of the bla(GES-2)-coding region distinguishes this ESBL from bla(GES-1) and the bla(IBC)-type genes, making it an ideal target for the development of a novel sequence-specific, peptide nucleic acid (PNA)-based multiplex PCR detection method. By using two primer pairs in conjunction with a PNA probe, this method provided an accurate means of identification of bla(GES-2) compared to standard PCR and gene sequencing techniques when it was used to test 100 P. aeruginosa clinical isolates as well as previously published, well-described control strains encompassing all presently known genes in the bla(GES-IBC) ESBL family. This novel method has the potential to be used in large-scale, cost-effective screening programs for specific or geographically restricted ESBLs.
Department of Medical Microbiology, Pathology, Faculty of Health Sciences, University of Pretoria, P.O. Box 2034, Pretoria 0001, South Africa. gweldhagen@medic.up.ac.za
Gerhard F
GF
Weldhagen
D003362:Cost-Benefit Analysis; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D020135:Peptide Nucleic Acids / Q000235:genetics*; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D018440:beta-Lactam Resistance; D001618:beta-Lactamases / Q000235:genetics*
D017931:DNA Primers; D004269:DNA, Bacterial; D020135:Peptide Nucleic Acids; C435769:beta-lactamase GES-2; D001618:beta-Lactamases