Characterization of microbial communities found in the human vagina by analysis of terminal restriction fragment length polymorphisms of 16S rRNA genes
TLDR
Findings show that T-RFLP of 16S rRNA genes can be used to compare vaginal microbial communities and gain information about the numerically dominant populations that are present, and are largely invariant over time.Abstract:
To define and monitor the structure of microbial communities found in the human vagina, a cultivation-independent approach based on analyses of terminal restriction fragment length polymorphisms (T-RFLP) of 16S rRNA genes was developed and validated. Sixteen bacterial strains commonly found in the human vagina were used to construct model communities that were subsequently used to develop efficient means for the isolation of genomic DNA and an optimal strategy for T-RFLP analyses. The various genera in the model community could best be resolved by digesting amplicons made using bacterial primers 8f and 926r with HaeIII; fewer strains could be resolved using other primer-enzyme combinations, and no combination successfully distinguished certain species of the same genus. To demonstrate the utility of the approach, samples from five women that had been collected over a 2-month period were analyzed. Differences and similarities among the vaginal microbial communities of the women were readily apparent. The T-RFLP data suggest that the communities of three women were dominated by a single phylotype, most likely species of Lactobacillus. In contrast, the communities of two other women included numerically abundant populations that differed from Lactobacillus strains whose 16S rRNA genes had been previously determined. The T-RFLP profiles of samples from all the women were largely invariant over time, indicating that the kinds and abundances of the numerically dominant populations were relatively stable throughout two menstrual cycles. These findings show that T-RFLP of 16S rRNA genes can be used to compare vaginal microbial communities and gain information about the numerically dominant populations that are present.read more
Citations
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Critical Evaluation of Two Primers Commonly Used for Amplification of Bacterial 16S rRNA Genes
Jeremy A. Frank,Claudia I. Reich,Shobha Sharma,Jon S. Weisbaum,Jon S. Weisbaum,Brenda A. Wilson,Gary J. Olsen +6 more
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Differences in the composition of vaginal microbial communities found in healthy Caucasian and black women
Xia Zhou,Celeste J. Brown,Zaid Abdo,Catherine C. Davis,Melanie A. Hansmann,Paul Joyce,James A. Foster,Larry J. Forney +7 more
TL;DR: It is postulate that because of differences in composition, not all vaginal communities are equally resilient, and that differences in the vaginal microbiota of Caucasian and black women may at least partly account for known disparities in the susceptibility of women in these racial groups to bacterial vaginosis and sexually transmitted diseases.
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Characterization of vaginal microbial communities in adult healthy women using cultivation-independent methods
Xia Zhou,Stephen J. Bent,Maria G. Schneider,Catherine C. Davis,Mohammed R. Islam,Larry J. Forney +5 more
TL;DR: Surprising results suggest that culture-independent methods can provide new insights into the diversity of bacterial species found in the human vagina, and this information could prove to be pivotal in understanding risk factors for various infectious diseases.
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The vaginal microbiome: New information about genital tract flora using molecular based techniques
Ronald F. Lamont,Jack D. Sobel,Robert A. Akins,Sonia S. Hassan,Tinnakorn Chaiworapongsa,Juan Pedro Kusanovic,Roberto Romero,Roberto Romero +7 more
TL;DR: This work presents new information about genital tract flora using molecular based techniques that helps clarify the role of “good bacteria” in women’s sex lives.
References
More filters
Journal ArticleDOI
Phylogenetic identification and in situ detection of individual microbial cells without cultivation.
TL;DR: Phylogenetic analysis of the retrieved rRNA sequence of an uncultured microorganism reveals its closest culturable relatives and may, together with information on the physicochemical conditions of its natural habitat, facilitate more directed cultivation attempts.
Book
Nucleic acid techniques in bacterial systematics
TL;DR: Isolation and purification of nucleic acids DNA reassociation experiments DNA-rRNA hybridization and methods DNA sequencing in bacterial systematics direct sequence analysis of small RNAs 16S/23S rRNA sequencing the polymerase chain reaction development and application of nucleics acid probes DNA fingerprinting from macromolecules to trees.
Journal ArticleDOI
The use of DAPI for identifying and counting aquatic microflora1
Karen G. Porter,Yvette S. Feig +1 more
TL;DR: Use of DAPI improved visualization and counting of <1-µm bacteria and blue-green algae in seston-rich samples and extended sample storage to at least 24 weeks.