Routine molecular identification of enterococci by gene-specific PCR and 16S ribosomal DNA sequencing.
For 279 clinically isolated specimens identified by commercial kits as enterococci, genotypic identification was performed by two multiplex PCRs, one with ddl(E. faecalis) and ddl(E. faecium) primers and another with vanC-1 and vanC-2/3 primers, and by 16S ribosomal DNA (rDNA) sequencing. For 253 strains, phenotypic and genotypic results were the same. Multiplex PCR allowed for the identification of 13 discordant results. Six strains were not enterococci and were identified by 16S rDNA sequencing. For 5 discordant and 10 concordant enterococcal strains, 16S rDNA sequencing was needed. Because many supplementary tests are frequently necessary for phenotypic identification, the molecular approach is a good alternative.
Angeletti
S
S
Libera Università Campus Bio-Medico, 00155 Rome, Italy.
Lorino
G
G
Gherardi
G
G
Battistoni
F
F
De Cesaris
M
M
Dicuonzo
G
G
D017931:DNA Primers; D004275:DNA, Ribosomal / Q000235:genetics*; D016983:Enterococcus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D013293:Enterococcus faecalis / Q000145:classification / Q000235:genetics; D016984:Enterococcus faecium / Q000145:classification / Q000235:genetics; D016908:Gram-Positive Bacterial Infections / Q000382:microbiology; D006801:Humans; D010453:Peptide Synthases / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D011933:Reagent Kits, Diagnostic; D010453:Peptide Synthases; C019638:D-alanylalanine synthetase