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Journal of Clinical Microbiology, October 2007, p. 3270-3276, Vol. 45, No. 10
0095-1137/07/$08.00+0     doi:10.1128/JCM.01272-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Targeted PCR for Detection of Vaginal Bacteria Associated with Bacterial Vaginosis{triangledown}

David N. Fredricks,1,2* Tina L. Fiedler,1 Katherine K. Thomas,3 Brian B. Oakley,1 and Jeanne M. Marrazzo2,3

Program in Infectious Diseases, Fred Hutchinson Cancer Research Center, Seattle, Washington,1 Division of Allergy and Infectious Diseases, Department of Medicine,2 Center for AIDS and STD, University of Washington, Seattle, Washington3

Received 24 June 2007/ Returned for modification 16 July 2007/ Accepted 30 July 2007

Several novel bacterial species have been detected in subjects with bacterial vaginosis (BV) by using broad-range PCR assays, but this approach is insensitive for detecting minority species. We developed a series of taxon-directed 16S rRNA gene PCR assays for more sensitive detection of key vaginal bacteria. We sought to determine the prevalence of each species in the vagina, its association with BV, and the utility of PCR for the microbiological diagnosis of BV. Targeted PCR assays were developed for 17 vaginal bacterial species and applied to 264 vaginal-fluid samples from 81 subjects with and 183 subjects without BV. The results were compared to those of two widely accepted methods for diagnosing BV, the use of clinical findings (Amsel criteria) and the interpretation of vaginal-fluid Gram stains (Nugent criteria). Leptotrichia/Sneathia, Atopobium vaginae, an Eggerthella-like bacterium, Megasphaera species, and three novel bacteria in the order Clostridiales are among the bacterial species significantly associated with BV. PCR detection of either a Megasphaera species or one of the Clostridiales bacteria yielded a sensitivity of 99% and a specificity of 89% for diagnosis of BV compared to the Amsel clinical criteria and a sensitivity of 95.9% and a specificity of 93.7% compared to the Nugent criteria (Gram stain). PCR detection of one or more fastidious bacterial species is a more reliable indicator of BV than detection of bacteria, such as Gardnerella vaginalis, previously linked to BV, highlighting the potential of PCR for the diagnosis of BV.

* Corresponding author. Mailing address: Program in Infectious Diseases, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue, North, D3-100, Box 19024, Seattle, WA 98109-1024. Phone: (206) 667-1935. Fax: (206) 667-4411. E-mail: dfredric{at}fhcrc.org

{triangledown} Published ahead of print on 8 August 2007.

Journal of Clinical Microbiology, October 2007, p. 3270-3276, Vol. 45, No. 10
0095-1137/07/$08.00+0     doi:10.1128/JCM.01272-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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