Background Bacterial vaginosis affects millions of women andis associated with several serious health conditions. The causeof bacterial vaginosis remains poorly understood despite numerousstudies based on cultures. Bacteria in microbial communitiescan be identified without cultivation by characterizing theirribosomal DNA (rDNA) sequences.
Methods We identified bacteria in samples of vaginal fluid witha combination of broad-range polymerase-chain-reaction (PCR)amplification of 16S rDNA with clone analysis, bacterium-specificPCR assay of 16S rDNA, and fluorescence in situ hybridization(FISH) performed directly on vaginal fluid from 27 subjectswith bacterial vaginosis and 46 without the condition. Twenty-onesubjects were studied with the use of broad-range PCR of 16SrDNA, and 73 subjects were studied with the use of bacterium-specificPCR.
Results Women without bacterial vaginosis had 1 to 6 vaginalbacterial species (phylotypes) in each sample (mean, 3.3), asdetected by broad-range PCR of 16S rDNA, and lactobacillus specieswere the predominant bacteria noted (83 to 100 percent of clones).Women with bacterial vaginosis had greater bacterial diversity(P<0.001), with 9 to 17 phylotypes (mean, 12.6) detectedper sample and newly recognized species present in 32 to 89percent of clones per sample library (mean, 58 percent). Thirty-fiveunique bacterial species were detected in the women with bacterialvaginosis, including several species with no close cultivatedrelatives. Bacterium-specific PCR assays showed that severalbacteria that had not been previously described were highlyprevalent in subjects with bacterial vaginosis but rare in healthycontrols. FISH confirmed that newly recognized bacteria detectedby PCR corresponded to specific bacterial morphotypes visiblein vaginal fluid.
Conclusions Women with bacterial vaginosis have complex vaginalinfections with many newly recognized species, including threebacteria in the Clostridiales order that were highly specificfor bacterial vaginosis.
Source Information
From the Program in Infectious Diseases, Fred Hutchinson Cancer Research Center (D.N.F., T.L.F.); and the Department of Medicine, Division of Allergy and Infectious Diseases, University of Washington (D.N.F., J.M.M.) both in Seattle.
Address reprint requests to Dr. Fredricks at the Program in Infectious Diseases, Fred Hutchinson Cancer Research Center, 1100 Fairview Ave. N., D3-100, Box 19024, Seattle, WA 98109-1024, or at dfredric{at}fhcrc.org.
Xu, J., Holzman, C. B., Arvidson, C. G., Chung, H., Goepfert, A. R.
(2008). Midpregnancy Vaginal Fluid Defensins, Bacterial Vaginosis, and Risk of Preterm Delivery. Obstet Gynecol
112: 524-531
[Abstract][Full Text]
Stoner, K. A., Rabe, L. K., Austin, M. N., Meyn, L. A., Hillier, S. L.
(2008). Quantitative Survival of Aerobic and Anaerobic Microorganisms in Port-A-Cul and Copan Transport Systems. J. Clin. Microbiol.
46: 2739-2744
[Abstract][Full Text]
Oakley, B. B., Fiedler, T. L., Marrazzo, J. M., Fredricks, D. N.
(2008). Diversity of Human Vaginal Bacterial Communities and Associations with Clinically Defined Bacterial Vaginosis. Appl. Environ. Microbiol.
74: 4898-4909
[Abstract][Full Text]
Brotman, R. M., Klebanoff, M. A., Nansel, T. R., Andrews, W. W., Schwebke, J. R., Zhang, J., Yu, K. F., Zenilman, J. M., Scharfstein, D. O.
(2008). A Longitudinal Study of Vaginal Douching and Bacterial Vaginosis--A Marginal Structural Modeling Analysis. Am J Epidemiol
168: 188-196
[Abstract][Full Text]
Marrazzo, J. M., Thomas, K. K., Fiedler, T. L., Ringwood, K., Fredricks, D. N.
(2008). Relationship of Specific Vaginal Bacteria and Bacterial Vaginosis Treatment Failure in Women Who Have Sex with Women. ANN INTERN MED
149: 20-28
[Abstract][Full Text]
Gelber, S. E., Aguilar, J. L., Lewis, K. L. T., Ratner, A. J.
(2008). Functional and Phylogenetic Characterization of Vaginolysin, the Human-Specific Cytolysin from Gardnerella vaginalis. J. Bacteriol.
190: 3896-3903
[Abstract][Full Text]
Frank, J. A., Reich, C. I., Sharma, S., Weisbaum, J. S., Wilson, B. A., Olsen, G. J.
(2008). Critical Evaluation of Two Primers Commonly Used for Amplification of Bacterial 16S rRNA Genes. Appl. Environ. Microbiol.
74: 2461-2470
[Abstract][Full Text]
Josey, W E, Schwebke, J R
(2008). The polymicrobial hypothesis of bacterial vaginosis causation: a reassessment. Int J STD AIDS
19: 152-154
[Abstract][Full Text]
Zozaya-Hinchliffe, M., Martin, D. H., Ferris, M. J.
(2008). Prevalence and Abundance of Uncultivated Megasphaera-Like Bacteria in the Human Vaginal Environment. Appl. Environ. Microbiol.
74: 1656-1659
[Abstract][Full Text]
Lawson, P. A., Moore, E., Falsen, E.
(2008). Prevotella amnii sp. nov., isolated from human amniotic fluid. Int. J. Syst. Evol. Microbiol.
58: 89-92
[Abstract][Full Text]
Marrazzo, J. M
(2007). Elusive aetiology of bacterial vaginosis. Do lesbians have a clue?. Sex. Transm. Infect.
83: 424-425
[Full Text]
Fredricks, D. N., Fiedler, T. L., Thomas, K. K., Oakley, B. B., Marrazzo, J. M.
(2007). Targeted PCR for Detection of Vaginal Bacteria Associated with Bacterial Vaginosis. J. Clin. Microbiol.
45: 3270-3276
[Abstract][Full Text]
Vitali, B., Pugliese, C., Biagi, E., Candela, M., Turroni, S., Bellen, G., Donders, G. G. G., Brigidi, P.
(2007). Dynamics of Vaginal Bacterial Communities in Women Developing Bacterial Vaginosis, Candidiasis, or No Infection, Analyzed by PCR-Denaturing Gradient Gel Electrophoresis and Real-Time PCR. Appl. Environ. Microbiol.
73: 5731-5741
[Abstract][Full Text]
Thilesen, C. M., Nicolaidis, M., Lokebo, J. E., Falsen, E., Jorde, A. T., Muller, F.
(2007). Leptotrichia amnionii, an Emerging Pathogen of the Female Urogenital Tract. J. Clin. Microbiol.
45: 2344-2347
[Abstract][Full Text]
Goto, M., Hitomi, S., Ishii, T.
(2007). Bacterial Arthritis Caused by Leptotrichia amnionii. J. Clin. Microbiol.
45: 2082-2083
[Abstract][Full Text]
Thies, F. L., Konig, W., Konig, B.
(2007). Rapid characterization of the normal and disturbed vaginal microbiota by application of 16S rRNA gene terminal RFLP fingerprinting. J Med Microbiol
56: 755-761
[Abstract][Full Text]
Ferris, M. J., Norori, J., Zozaya-Hinchliffe, M., Martin, D. H.
(2007). Cultivation-Independent Analysis of Changes in Bacterial Vaginosis Flora Following Metronidazole Treatment. J. Clin. Microbiol.
45: 1016-1018
[Abstract][Full Text]
Eckert, L. O.
(2006). Acute Vulvovaginitis. NEJM
355: 1244-1252
[Full Text]
Baron, R. M., Baron, M. J., Perrella, M. A.
(2006). Pathobiology of Sepsis: Are We Still Asking the Same Questions?. Am. J. Respir. Cell Mol. Bio.
34: 129-134
[Full Text]
Hale, L. P., Swidsinski, A., Mendling, W., Fredricks, D. N., Marrazzo, J. M.
(2006). Bacteria Associated with Bacterial Vaginosis. NEJM
354: 202-203
[Full Text]
(2005). Bacterial Vaginosis: New Bugs Revealed. JWatch Women's Health
2005: 3-3
[Full Text]
(2005). New Insights into Bacterial Vaginosis. JWatch Infect. Diseases
2005: 4-4
[Full Text]