Development of the Specific and Random Amplification (SARA)-PCR for both species identification of enterococci and detection of the vanA gene

• Published in: Journal of microbiological methods 2001, Vol.43 (3), p.233-239
• Main Authors: Knijff, Edo, Dellaglio, Franco, Lombardi, Angiolella, Biesterveld, Steef, Torriani, Sandra
• Format: Article
• Language: English
• Subjects: Animals; Anti-Bacterial Agents - pharmacology; Bacterial Proteins - genetics; Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; Carbon-Oxygen Ligases - genetics; Enterococci; Enterococcus; Enterococcus - classification; Enterococcus - drug effects; Enterococcus - genetics; Food Microbiology; Fundamental and applied biological sciences. Psychology; Gene Amplification; Genes, Bacterial; Gram-Positive Bacterial Infections - microbiology; Humans; Identification; Microbial Sensitivity Tests; Microbiology; Polymerase Chain Reaction - methods; SARA-PCR; Species Specificity; Teicoplanin - pharmacology; vanA gene; vanA gene detection; Vancomycin - pharmacology; Vancomycin Resistance
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/S0167-7012(00)00225-6

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 ∼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.