Detection of vancomycin resistance genes combined with typing of Enterococci by means of multiplex PCR amplification and multiple primer DNA sequencing.
A multiplex PCR assay for the detection of vancomycin resistance (van) genes in enterococci was established. Primers targeting the 16S rRNA gene were included in the reaction mixture. Multiple-primer DNA sequencing of the PCR products provided species identification through partial nucleotide sequences of 16S rRNA genes, as well as confirmation of the correct identification of vanA, vanB, vanC-1, and vanC-2/3 genotypes. Thirty-nine enterococcal clinical isolates and type strains were examined for the presence of vancomycin resistance determinants. Twelve other isolates from a clinical reference collection (some of them having vanA, vanB, vanC-1, or vanC-2/3 genotypes) were used as controls. Hybridization and partial DNA sequence analysis of multiplex PCR products revealed that none of the clinical isolates had a vanA genotype and only one had a vanB genotype. vanC-1 was found in three clinical isolates, and vanC-2/3 in one. Results obtained with the reference and type strains were in agreement with earlier results.
Monstein
H J
HJ
Division of Clinical Microbiology, Molecular Biology Laboratory-LMO, Faculty of Health Sciences, University Hospital, Linköping, Sweden.
Johansson
Y
Y
Jonasson
J
J
D015373:Bacterial Typing Techniques; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D016983:Enterococcus / Q000145:classification* / Q000187:drug effects / Q000235:genetics*; D005798:Genes, Bacterial; D005838:Genotype; D006801:Humans; D008826:Microbial Sensitivity Tests; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods*; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D013045:Species Specificity; D020713:Vancomycin Resistance / Q000235:genetics*
D017931:DNA Primers; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S