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Enterococci are nosocomial pathogens that can form biofilms, which contribute to their virulence and antibiotic resistance. Although many genes involved in biofilm formation have been defined, their distribution among enterococci has not been comprehensively studied on a genome scale, and their diagnostic ability to predict biofilm phenotypes is not fully established. Here, we assessed the biofilm-forming ability of 90 enterococcal clinical isolates. Major patterns of virulence gene distribution in enterococcal genomes were identified, and the differentiating virulence genes were screened by polymerase chain reaction (PCR) in 31 of the clinical isolates. We found that detection of  gelE  in  Enterococcus faecalis  is not sufficient to predict gelatinase activity unless  fsrAB , or  fsrB  alone, is PCR-positive ( P =  0.0026 and 0.0012, respectively). We also found that  agg  is significantly enriched in isolates with medium and strong biofilm formation ability ( P =  0.0026). Additionally, vancomycin, applied at sub minimal inhibitory concentrations, inhibited biofilm in four out of five strong biofilm-forming isolates. In conclusion, we suggest using  agg  and  fsrB  genes, together with the previously established  gelE , for better prediction of biofilm strength and gelatinase activity, respectively. Future studies should explore the mechanism of biofilm inhibition by vancomycin and its possible use for antivirulence therapy.

Biofilm formation in enterococci: genotype-phenotype correlations and inhibition by vancomycin