Development of the Specific and Random Amplification (SARA)-PCR for both species identification of enterococci and detection of the vanA gene.
A rapid and reliable polymerase chain reaction (Specific and Random Amplification (SARA)-PCR) has been developed to identify enterococcal species and to detect the vanA gene in one single reaction. This technique was based on the use of the primer set previously designed to amplify a part of the vanA gene (Dutka-Malen et al., J. Clin. Microbiol., 33 (1995) 24-27). In the chosen low stringency conditions complex patterns were obtained, with a sharp band of approximately 700 bp in cases where the vanA gene was present. Discrimination at the species and strain level was achieved by applying the SARA-PCR assay to a collection of 55 enterococcal isolates and type strains. Simultaneously the vanA gene was detected in all strains showing high resistance to vancomycin.
Knijff
E
E
Dipartimento Scientifico e Tecnologico, Università degli Studi di Verona, Strada le Grazie 15, 37134, Verona, Italy.
Dellaglio
F
F
Lombardi
A
A
Biesterveld
S
S
Torriani
S
S
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics*; D019729:Carbon-Oxygen Ligases / Q000235:genetics*; D016983:Enterococcus / Q000145:classification* / Q000187:drug effects / Q000235:genetics*; D005516:Food Microbiology; D005784:Gene Amplification; D005798:Genes, Bacterial; D016908:Gram-Positive Bacterial Infections / Q000382:microbiology*; D006801:Humans; D008826:Microbial Sensitivity Tests; D016133:Polymerase Chain Reaction / Q000379:methods*; D013045:Species Specificity; D017334:Teicoplanin / Q000494:pharmacology; D014640:Vancomycin / Q000494:pharmacology; D020713:Vancomycin Resistance
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; C071437:VanA ligase, Bacteria; D017334:Teicoplanin; D014640:Vancomycin; D019729:Carbon-Oxygen Ligases