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Journal of Clinical Microbiology, July 2003, p. 3260-3264, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.3260-3264.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Optimization of Media for Detection of Hydrogen Peroxide Production by Lactobacillus Species

L. K. Rabe1 and S. L. Hillier1,2*

Magee-Women's Research Institute,1 Department of Obstetrics, Gynecology, and Reproductive Sciences, The University of Pittsburgh, Pittsburgh, Pennsylvania 152132

Received 24 October 2002/ Accepted 31 October 2002

A healthy vaginal ecosystem has been shown to be protective against the acquisition of human immunodeficiency virus and gonorrhea, and women who are colonized with H2O2-producing lactobacilli are more likely to maintain a normal vaginal flora than women with lactobacilli that do not produce H2O2. The purpose of this study was to formulate a testing medium that better supports the growth and detection of H2O2 by a broader range of lactobacilli than a published, widely used agar formulation (TMB). The new medium (TMB-Plus) consists of brucella agar base, 3,3',5,5'-tetramethylbenzidine, horseradish peroxidase, starch, vitamin K, hemin, magnesium sulfate, manganese sulfate, and horse serum. To validate the new formula, 256 vaginal isolates and ATCC strains were inoculated onto TMB-Plus and, for comparison, onto TMB. Growth was enhanced for 69% of the isolates on TMB-Plus, and 48% had enhanced color production. The percentage of H2O2-positive isolates increased from 71% on TMB to 79% on TMB-Plus. Formulations using Rogosa or MRS agar base in combination with peroxidase and a chromogen did not support the growth of all of the strains of Lactobacillus, and fewer H2O2-producing strains were detected on these formulations than on TMB-Plus. This new medium better supports the growth of a wider range of Lactobacillus strains isolated from the vagina and enhances the color production of H2O2-producing strains.

* Corresponding author. Mailing address: University of Pittsburgh, Department of Obstetrics, Gynecology, and Reproductive Sciences, Magee-Women's Hospital, 300 Halket St., Pittsburgh, PA 15213-3180. Phone: (412) 641-6416. Fax: (412) 641-1113. E-mail: slh6{at}pitt.edu.

Journal of Clinical Microbiology, July 2003, p. 3260-3264, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.3260-3264.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Boyd, M. A., Antonio, M. A. D., Hillier, S. L. (2005). Comparison of API 50 CH Strips to Whole-Chromosomal DNA Probes for Identification of Lactobacillus Species. J. Clin. Microbiol. 43: 5309-5311 [Abstract] [Full Text]  
  • Austin, M. N., Beigi, R. H., Meyn, L. A., Hillier, S. L. (2005). Microbiologic Response to Treatment of Bacterial Vaginosis with Topical Clindamycin or Metronidazole. J. Clin. Microbiol. 43: 4492-4497 [Abstract] [Full Text]  


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