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Journal of Clinical Microbiology, July 2001, p. 2439-2444, Vol. 39, No. 7
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.7.2439-2444.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Evaluation of Current Methods for Detection of Staphylococci with Reduced Susceptibility to Glycopeptides

Timothy R. Walsh,1,* Anne Bolmström,2 Anette Qwärnström,2 Phion Ho,2 Mandy Wootton,3 Robin A. Howe,3 Alasdair P. MacGowan,3 and Dan Diekema4

Department of Pathology and Microbiology, School of Medical Sciences, University of Bristol, Bristol BS8 1TD,1 and Bristol Centre for Antimicrobial Research and Evaluation, North Bristol NHS Trust and University of Bristol, and Department of Microbiology, Southmead Hospital, Bristol BS10 5NB,3 United Kingdom; AB BIODISK Research Laboratories, Dalvägen 10, S-169 56 Solna, Sweden2; and Anti-Infectives Research Center, Iowa City, Iowa4

Received 10 November 2000/Returned for modification 14 January 2001/Accepted 12 April 2001

The sensitivity and specificity of seven methods (agar dilution, broth microdilution, Etest at 0.5 and 2.0 McFarland (McF) inocula, two agar screening methods, and population studies [PS]) were evaluated in a double-blind study involving 284 methicillin-resistant Staphylococcus aureus (MRSA) strains and 45 Staphylococcus strains with reduced susceptibilities to vancomycin (SRSV). The results were compared to the population analysis profile-area under the curve ratio method (PAP-AUC ratio compared to that of Mu3) as described by Wootton et al. The agar screening method using brain heart infusion agar (6 µg of vancomycin per ml) gave a sensitivity of 22% and a specificity of 97%. A similar method using Mueller-Hinton agar (5 µg of vancomycin per ml) gave a sensitivity of 20% and a specificity of 99%. The PS method detected 34 false positives (12%) and gave a sensitivity of 71% and a specificity of 88%. Etest using 0.5 and 2.0 McF inocula gave sensitivities and specificities of 82 and 93% and of 96 and 97%, respectively. The best Etest interpretative criteria for the 2.0 McF inoculum was >= 8 mg of vancomycin per liter and >= 8 µg teicoplanin per ml or >= 12 µg of teicoplanin per ml. The direct colony suspension inoculum for this method was found to be equally accurate in detecting (hetero-)glycopeptide-intermediate S. aureus compared to the overnight broth inoculum preparation method. Agar dilution and broth microdilution using the NCCLS breakpoint criteria for vancomycin gave sensitivities and specificities of 20 and 100% and of 11 and 100%, respectively. Using the Etest with a 2.0 McF inoculum, six different media were assessed against a selection of SRSV (n = 48) and MRSA (n = 12). Brain heart infusion agar yielded the highest sensitivity and specificity values: 88 and 88%, respectively.

* Corresponding author. Mailing address: Department of Pathology and Microbiology, University of Bristol, Bristol BS8 1TD, United Kingdom. Phone: 44 (0)117-9287897. Fax: 44 (0)117-9287896. E-mail: t.r.walsh{at}bristol.ac.uk.

Journal of Clinical Microbiology, July 2001, p. 2439-2444, Vol. 39, No. 7
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.7.2439-2444.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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