title: CPT-EIA assays for the detection of vancomycin resistant vanA and vanB genes in enterococci.
abstract: Cycling Probe Technology (CPT) was combined with a colorimetric enzyme-immuno assay (EIA) to develop two assays for the detection of vanA and vanB genes in vancomycin resistant enterococci (VRE). The CPT-EIA assay employs a gene-specific fluorescein labeled DNA-RNA-DNA probe that gets cleaved within the probe : target duplex. The cleaved DNA probe fragments dissociate from the target, making it available for further cycling. Following the separation of cleaved probe fragments, anti-fluorescein-horseradish peroxidase antibodies are used for the detection of uncleaved probes. The two CPT-EIA assays were used to screen a collection of 440 clinical isolates (Modrusan et al., 1999). All of the 154 VanA and 131 VanB isolates were correctly identified in the vanA and vanB CPT-EIA, respectively. The VanA and VanB isolates were differentiated from vancomycin sensitive enterococci (VSE) and also from the VanC isolates. In addition, an accurate VRE detection in the CPT-EIA assay was shown with cultures grown on eight different media.
authors.affiliation: Incyte Pharmaceuticals, Microarray systems, Fremont, CA 94555, USA. zoram@incyte.com
authors.forename: Z
authors.initials: Z
authors.lastname: Modrusan
authors.forename: C
authors.initials: C
authors.lastname: Marlowe
authors.forename: D
authors.initials: D
authors.lastname: Wheeler
authors.forename: M
authors.initials: M
authors.lastname: Pirseyedi
authors.forename: R N
authors.initials: RN
authors.lastname: Bryan
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics*; D019729:Carbon-Oxygen Ligases / Q000235:genetics*; D003470:Culture Media; D013293:Enterococcus faecalis / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D016984:Enterococcus faecium / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D005798:Genes, Bacterial; D006801:Humans; D007124:Immunoenzyme Techniques; D014640:Vancomycin / Q000494:pharmacology*; D020713:Vancomycin Resistance
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D003470:Culture Media; C071437:VanA ligase, Bacteria; C079913:VanB protein, Enterococcus; D014640:Vancomycin; D019729:Carbon-Oxygen Ligases