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Try out PMC Labs and tell us what you think. Learn More. Species of the Lactobacillus acidophilus complex are generally considered to constitute most of the vaginal Lactobacillus flora, but the flora varies between studies. However, this may be due to difficulties in identifying the closely related species within the L. Two hundred two isolates from the vaginal fluids of 23 Swedish women without bacterial vaginosis, as defined by the criteria of Nugent et al.

Nugent, M. Krohn, and S. Hillier, J. The vaginal flora of most participants was dominated by a single RAPD type, but five of them harbored two RAPD types representing two different species or strains. The most frequently occurring species were Lactobacillus crispatusLactobacillus gasseriLactobacillus inersand Lactobacillus jensenii.

The healthy human vagina is dominated by lactobacilli, which play an important role in protecting the host from urogenital infections 417 Furthermore, it is widely recognized that the microbial balance between lactobacilli as the dominating flora and other, mainly gram-negative anaerobes can be upset and frequently result in Free Brescia ca pussy syndrome of bacterial vaginosis Thus, the microbial status of the healthy human vagina has to be defined in any study of vaginal lactobacilli.

The clinical criteria of Amsel et al. These scores are generally considered to be useful in treating patients with bacterial vaginosis but can also be used to define a healthy vaginal status by excluding bacterial vaginosis and by combining the with a short medical history and examination for sexually transmitted diseases. A general opinion is that species of the Lactobacillus acidophilus complex constitute most of the healthy vaginal Lactobacillus flora However, other species have been encountered, e. The group of organisms ly known as L.

The species in the complex are obligately homofermentative and belong to the rRNA group L. The closely related species within the L. In addition, the lack of reliable identification methods may for some of the variation in the Lactobacillus flora of the vagina found in different studies 23 Consequently, accurate genomic methods are needed in order to define the composition of the Lactobacillus flora in the vagina, not only for the treatment of infectious diseases but also to establish the normal Lactobacillus flora in different settings. With this objective, we applied PCR-based methods for the identification Free Brescia ca pussy the Lactobacillus flora.

Two hundred two isolates from the healthy vaginas of 23 Swedish women were typed and identified. As a key for the identification, 26 type and reference strains were used: L. Most women came for their scheduled cervical screening. Samples were collected from the vaginal fluid with a cotton swab rolled over the upper third part of the lateral vaginal wall.

The swabs were placed in modified Stuart medium Copan transport medium; Venturi Transystem, Brescia, Italy and sent to the laboratory, where they were cultured on the agar plates within 3 h. Another swab was similarly rolled over the same vaginal area and onto a glass slide. The slides were air dried, fixed by heat, and Gram stained. The slides were then assessed according to the criteria of Nugent et al.

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The specimens were vortexed for 5 s in 1 ml of phosphate-buffered saline pH 7. Lactobacilli from 23 women with normal vaginal flora according to the criteria of Nugent et al. One hundred eighty-six isolates with the ability to grow on Rogosa agar plates were randomly selected and studied, together with 16 isolates that grew only on horse blood agar. This template was used for all of the PCR-based analyses described below. Two hundred two isolates and the type strains were subjected to RAPD analysis. PCR amplification and agarose gel electrophoresis were performed as follows, as ly described by Quednau et al.

The gel images were then analyzed and grouped by GelCompar 4. In total, 72 vaginal isolates and all type and reference strains were subjected to TTGE. A standard composed of TTGE products of strains that showed clear and bright bands at different positions was used to allow gel-to-gel comparison. After this, the gel images were analyzed by GelCompar 4. Twenty-six vaginal isolates and all of the reference and type strains were assessed by multiplex PCR as ly described by Song et al.

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The other modification to the protocol was that 1. The sequencing reaction products were purified by applying streptavidin-coated superparamagnetic be Dynabe M Streptavidin; Dynal Biotech, Oslo, Norway prior to loading the sequencing gel. No single subject was dominated by more than two RAPD types. In a majority of the individuals, only one RAPD type was found. Exceptions were the patterns for L. TTGE patterns of representative vaginal Lactobacillus strains. Lanes S, standard. Lanes 1 to 6, L. Lanes 7 to 11, L. Lane 12, L. The multiplex PCR enabled the identification to the species level of vaginal isolates belonging to L.

Using the combined of the different methods, the complete identification showed that L. The predominant species of Lactobacillus found in the vaginas of healthy women as determined by reliable laboratory criteria those of Nugent et al.

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RAPD analysis was used in this study to group the isolates and to observe the heterogeneity of the Lactobacillus flora of each subject. RAPD analysis has ly been used to differentiate separate strains of Lactobacillus spp. Nevertheless, these isolates could be identified as L.

The sequence data on Free Brescia ca pussy vaginal isolates of L. In addition to the finding of L. Antonio et al. Lachlak et al. To our knowledge, L. One explanation for the absence of L. Species such as L.

None of them were found in the present study, and neither was the recently described species Lactobacillus fornicaliswhich was isolated from the posterior fornix of the human vagina 5. The differences in Lactobacillus flora between studies may be attributed to a of factors.

One suggestion is that the intestinal lactobacilli differ in western countries and Japan 26and the same may be true for the vagina However, the difference in the Lactobacillus flora is unlikely to represent geographical variation alone. More likely explanations are variations in the way that samples are taken and treated, the vaginal status, or the fact that identification has often been based on phenotypic methods 18 Phenotypic criteria are particularly unreliable for identification of members of the L.

National Center for Biotechnology InformationU. Journal List J Clin Microbiol v. J Clin Microbiol. Author information Article notes Copyright and information Disclaimer. Mailing address: Food Technology, P. BoxSE 00 Lund, Sweden. Phone: Fax: E-mail: es. This article has been cited by other articles in PMC. Abstract Species of the Lactobacillus acidophilus complex are generally considered to constitute most of the vaginal Lactobacillus flora, but the flora varies between studies. RAPD analysis. Multiplex PCR. TABLE 1. Vaginal isolates derived from 23 women by the different genotypic approaches.

Open in a separate window. Amsel, R. Totten, C. Spiegel, K. Chen, D. Eschenbach, and K. Nonspecific vaginitis. Diagnostic criteria and microbial and epidemiologic associations. Antonio, M. Hawes, and S. The identification of vaginal Lactobacillus species and the demographic and microbiologic characteristics of women colonized by these species.

Bohak, I. Back, L. Richter, M. Ehrmann, W. Ludwig, and K. Lactobacillus amylolyticus sp. Boris, S. Adherence of human vaginal lactobacilli to vaginal epithelial cells and interaction with uropathogens. Dicks, L. Silvester, P. Lawson, and M. Lactobacillus fornicalis sp. E vol. Efthymiou, C.

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