Open AccessSequence-Selective Recognition of Extended-Spectrum β-Lactamase GES-2 by a Competitive, Peptide Nucleic Acid-Based Multiplex PCR Assay
Author(s) -
Gerhard F. Weldhagen
Publication year - 2004
Publication title -
antimicrobial agents and chemotherapy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.07
H-Index - 259
eISSN - 1070-6283
pISSN - 0066-4804
DOI - 10.1128/aac.48.9.3402-3406.2004
Subject(s) - biology , multiplex polymerase chain reaction , peptide nucleic acid , nucleic acid , gene , nucleic acid sequence , multiplex , primer (cosmetics) , imipenem , pseudomonas aeruginosa , computational biology , polymerase chain reaction , microbiology and biotechnology , genetics , bacteria , chemistry , organic chemistry
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.