Convenient selective differential broth for isolation of vancomycin-resistant enterococcus from fecal material.
Studies have shown that vancomycin broth enrichment is superior to direct plating for the detection of vancomycin-resistant enterococcus (VRE), but vancomycin selective broth is not generally commercially available. We developed an easy-to-prepare VRE selective differential broth and compared it to direct plating on bile esculin azide (BEA) agar for the isolation of VRE from fecal samples. A total of 528 consecutive rectal swabs and stools were inoculated onto BEA agar and into BEA broth with vancomycin at a concentration of 15 microg/ml (BEA VAN15 microg/ml broth). After 1 to 2 days of incubation, broths were subcultured to BEA VAN6 microg/ml agar. Bile esculin-positive colonies from the direct and broth subculture plates were evaluated for the presence of VRE by standard microbiological techniques. Addition of the broth enrichment step led to the detection of significantly more VRE isolates than did direct plating alone (28 versus 18 VRE isolates, respectively). In all, 30 VRE strains were isolated from 29 cultures, all of which were Enterococcus faecium. MICs of vancomycin ranged from 32 microg/ml (n = 2) to > 256 microg/ml (n = 28). Twenty-two VRE isolates were available for further testing: sixteen exhibited a VanA phenotype and six were of the VanB phenotype. van genotypes were in agreement with phenotypes for all VRE isolates except one, which could not be genotyped. The broth method also resulted in significantly fewer bile esculin-positive, non-VRE isolates requiring further workup. We have thus developed an easily prepared vancomycin selective differential broth that is significantly more sensitive and specific in the detection of VRE than is direct fecal plating to BEA agar.
Novicki
Thomas J
TJ
Department of Laboratory Medicine, University of Washington, Seattle, Washington 98195-7110, USA.
Schapiro
Jeffrey M
JM
Ulness
Bruce K
BK
Sebeste
Ann
A
Busse-Johnston
Laurel
L
Swanson
Kristine M
KM
Swanzy
Susan R
SR
Leisenring
Wendy
W
Limaye
Ajit P
AP
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001431:Bacteriological Techniques / Q000379:methods*; D003470:Culture Media / Q000737:chemistry; D016984:Enterococcus faecium / Q000145:classification / Q000187:drug effects* / Q000302:isolation & purification*; D005243:Feces / Q000382:microbiology*; D016908:Gram-Positive Bacterial Infections / Q000382:microbiology; D006801:Humans; D008826:Microbial Sensitivity Tests; D012680:Sensitivity and Specificity; D014640:Vancomycin / Q000494:pharmacology; D020713:Vancomycin Resistance
D000900:Anti-Bacterial Agents; D003470:Culture Media; D014640:Vancomycin