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Journal of Clinical Microbiology, April 2004, p. 1637-1640, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1637-1640.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Convenient Selective Differential Broth for Isolation of Vancomycin-Resistant Enterococcus from Fecal Material

Thomas J. Novicki,1,{dagger} Jeffrey M. Schapiro,1,2 Bruce K. Ulness,1 Ann Sebeste,1 Laurel Busse-Johnston,1 Kristine M. Swanson,1 Susan R. Swanzy,1 Wendy Leisenring,3 and Ajit P. Limaye1,2*

Departments of Laboratory Medicine,1 Medicine, University of Washington,2 Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, Washington3

Received 31 October 2003/ Returned for modification 24 November 2003/ Accepted 19 December 2003

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 µg/ml (BEA VAN15µg/ml broth). After 1 to 2 days of incubation, broths were subcultured to BEA VAN6µg/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 µg/ml (n = 2) to > 256 µg/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.

* Corresponding author. Mailing address: University of Washington, Dept. of Laboratory Medicine, 1959 N.E. Pacific St., NW120, Seattle, WA 98195-7110. Phone: (206) 598-6131. Fax: (206) 598-6189. E-mail: limaye{at}u.washington.edu.

{dagger} Present address: Microbiology Section, Marshfield Laboratories, Marshfield, WI 54449.

Journal of Clinical Microbiology, April 2004, p. 1637-1640, Vol. 42, No. 4
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.4.1637-1640.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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