CPT-EIA assays for the detection of vancomycin resistant vanA and vanB genes in enterococci.
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.
Modrusan
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Z
Incyte Pharmaceuticals, Microarray systems, Fremont, CA 94555, USA. zoram@incyte.com
Marlowe
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C
Wheeler
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D
Pirseyedi
M
M
Bryan
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RN
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
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D003470:Culture Media; C071437:VanA ligase, Bacteria; C079913:VanB protein, Enterococcus; D014640:Vancomycin; D019729:Carbon-Oxygen Ligases