CPT-EIA assays for the detection of vancomycin resistant vanA and vanB genes in enterococci
| Autor: | Robert N. Bryan, Donald Wheeler, Mona Pirseyedi, Zora Modrusan, Cherith Marlowe |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Microbiology (medical)
Enterococcus faecium Microbiology Immunoenzyme Techniques chemistry.chemical_compound Bacterial Proteins Vancomycin Vancomycin resistant medicine Enterococcus faecalis Humans Fluorescein Gene Carbon-Oxygen Ligases biology Hybridization probe Vancomycin Resistance General Medicine biochemical phenomena metabolism and nutrition bacterial infections and mycoses biology.organism_classification Streptococcaceae Molecular biology Anti-Bacterial Agents Culture Media carbohydrates (lipids) Infectious Diseases chemistry Enterococcus Genes Bacterial Bacteria medicine.drug |
| Zdroj: | Diagnostic microbiology and infectious disease. 37(1) |
| ISSN: | 0732-8893 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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