title: vanA-targeted oligonucleotide DNA probe designed to monitor vancomycin- and teicoplanin-resistant bacteria in surface waters.
abstract: The glycopeptide antibiotics teicoplanin and vancomycin are common to treat severe Gram-positive bacterial infections. The gene vanA confers high-level resistance to these antibiotics, and these phenomena have been shown to be transferable. Release of vancomycin- and teicoplanin-resistant bacteria to surface waters is, therefore, of particular concern since they might proliferate and spread in different environments. Monitoring of the fate of vanA gene in these waters provides information on the exposure and potential threats of those bacteria for the environment and public health. Therefore, this study aimed at preparing a 25-mer-oligonucleotide DNA probe based on the 909 bp BamHI-ClaI fragment from Enterococcus faecium plasmids pVEF1 and pVEF2 through the use of Vector NTI Express Software. The newly designed vanA probe displayed highly specific hybridization with vanA-positive Enterococcus faecalis tested at 46 °C, 55 % formamide, and 0.020 M NaCl stringency conditions. In situ fluorescein hybridizations under the same stringency conditions were also used to monitor river water samples by using fluorescein microscopy. The results showed that the vanA-targeted oligonucleotide DNA probe prepared was not only highly specific but also quantitative tool for monitoring vancomycin- and teicoplanin-resistant bacteria in surface waters.
authors.affiliation: Department of Biotechnology, Middle East Technical University, 06800, Ankara, Turkey.
authors.forename: Mustafa
authors.initials: M
authors.lastname: Nakipoglu
authors.affiliation: Department of Environmental Engineering, Middle East Technical University, 06800, Ankara, Turkey.
authors.forename: Fadime
authors.initials: F
authors.lastname: Yilmaz
authors.affiliation: Department of Biotechnology, Middle East Technical University, 06800, Ankara, Turkey. bicgen@metu.edu.tr.|Department of Environmental Engineering, Middle East Technical University, 06800, Ankara, Turkey. bicgen@metu.edu.tr.
authors.forename: Bulent
authors.initials: B
authors.lastname: Icgen
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics*; D019729:Carbon-Oxygen Ligases / Q000235:genetics*; D015342:DNA Probes; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics; D013293:Enterococcus faecalis / Q000187:drug effects* / Q000235:genetics; D016984:Enterococcus faecium / Q000187:drug effects* / Q000235:genetics; D004784:Environmental Monitoring / Q000379:methods*; D016908:Gram-Positive Bacterial Infections / Q000188:drug therapy; D006801:Humans; D009841:Oligonucleotides; D017334:Teicoplanin / Q000494:pharmacology*; D014640:Vancomycin / Q000494:pharmacology*; D014871:Water Microbiology
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D015342:DNA Probes; D009841:Oligonucleotides; C071437:VanA ligase, Bacteria; D017334:Teicoplanin; D014640:Vancomycin; D019729:Carbon-Oxygen Ligases
keywords: Glycopeptide resistance; Surface waters; Teicoplanin; Vancomycin; vanA DNA probe