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Lactobacilli dominating the normal vaginal flora
Linköping University, Department of Molecular and Clinical Medicine, Clinical Microbiology. Linköping University, Department of Molecular and Clinical Medicine, Obstetrics and gynecology. Linköping University, Faculty of Health Sciences.
2003 (English)Licentiate thesis, comprehensive summary (Other academic)
Abstract [en]

The microbiology of the ecological niche of the vagina is dynamic. There are numerous inhabitants, mainly anaerobic bacteria. During the fertile years the vaginal mucosa is normally dominated by lactobacilli, a fact that has been known for over a century. Lactobacilli are a phenotypically heterogeneous group of bacteria that first became possible to identify to the species level with some precision using recently developed nucleic acid based techniques. In this study vaginal fluid was cultured from women scheduled for their regular PAP smear. Two hundred and two isolates from 23 women with normal vaginal fluid were typed by randomly amplified polymorphic DNA (RAPD) PCR analysis and identified to the species level by temporal temperature gradient gel electrophoresis (TTGE), and 16S rDNA sequencing. Four hundred and four isolates from 23 women were typed with broad range PCR of 16S rRNA gene region V I and V3 by pyrosequencing. Most women harboured one single species, a few had two different species, and only one woman harboured more than two different species of lactobacilli. The species that were found, which were similar in the two studies, were: Lactobacillus crispalus, L. gasseri, L. jensenii and L. iners. L. iners has never been reported before as a member of the dominant normal vaginal lactobacillary flora. Under some conditions, which have not been determined with certainty, the lactobacilli are overgrown by large amounts of Gardnerella vaginalis and anaerobic bacteria, mainly Bacleroides spp and Mobiluncus, causing the syndrome of Bacterial Vaginosis (BV). BV is associated with several severe reproductive and genitourinary complications in women. A major issue in studying normal Lactobacilli in vaginal fluid samples from women of reproductive age is to differentiate the normal flora from that of BV. To sharpen the diagnostic tools (i.e. Nugent scoring of Gram stained slides) for separation of normal flora from that of BV, a workshop was set up. The major results showed discrepancies in diagnosis when there were very few lactobacilli. There was disagreement on where to delimit small lactobacilli from G. vaginalis and Bacteroides. The use of scoring to delimit normal flora, as was done in the workshop, is proposed as a prerequisite for further studies of normal vaginal flora

Place, publisher, year, edition, pages
Linköping: Linköpings universitet , 2003. , 21 p.
Series
Linköping Studies in Health Sciences. Thesis, ISSN 1100-6013 ; 61
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:liu:diva-26680Local ID: 11247ISBN: 91-7373-544-2 (print)OAI: oai:DiVA.org:liu-26680DiVA: diva2:247229
Presentation
2003-05-15, Flemingsalen, Universitetssjukhuset, Linköping, 13:00 (Swedish)
Available from: 2009-10-08 Created: 2009-10-08 Last updated: 2013-09-12
List of papers
1. Vaginal Lactobacillus flora of healthy Swedish women
Open this publication in new window or tab >>Vaginal Lactobacillus flora of healthy Swedish women
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2002 (English)In: Journal of clinical microbiology, ISSN 0095-1137 (print) 1098-660X (online), Vol. 40, no 8, 2746-2749 p.Article in journal (Refereed) Published
Abstract [en]

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. acidophilus complex by using traditional methods and to variations in the vaginal status of the participants. Two hundred two isolates from the vaginal fluids of 23 Swedish women without bacterial vaginosis, as defined by the criteria of Nugent et al. (R. P. Nugent, M. A. Krohn, and S. L. Hillier, J. Clin. Microbiol. 29:297-301, 1991), were typed by randomly amplified polymorphic DNA (RAPD) analysis and identified to the species level by temporal temperature gradient gel electrophoresis, multiplex PCR, and 16S ribosomal DNA sequencing. 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 crispatus, Lactobacillus gasseri, Lactobacillus iners, and Lactobacillus jensenii. L. iners has not previously been reported as one of the predominant Lactobacillus species in the vagina.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-13051 (URN)10.1128/JCM.40.8.2746-2749.2002 (DOI)
Available from: 2008-03-19 Created: 2008-03-19 Last updated: 2013-09-12
2. Identification of randomly selected colonies of Lactobacilli from normal vaginal fluid by pyrosequencing of the 16S rDNA Variable V1 and V3 Regions
Open this publication in new window or tab >>Identification of randomly selected colonies of Lactobacilli from normal vaginal fluid by pyrosequencing of the 16S rDNA Variable V1 and V3 Regions
2002 (English)In: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 110, no 11, 802-810 p.Article in journal (Refereed) Published
Abstract [en]

The present study aimed to characterize lactobacilli in vaginal fluid from 23 adult healthy women by using high-throughput DNA sequencing for identification of a large number of randomly selected colonies appearing on Rogosa and blood agar. The typing method was based on broad-range PCR of 16S rRNA gene variable regions V1 and V3, pyrosequencing, and classification of the fragments by alignment with NCBI-catalogued sequences and type strain sequences. Four major groups of sequences were found among the 402 isolates clearly corresponding to Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus iners and Lactobacillus jensenii when compared to the sequences obtained for type strains. Our results indicate that pyrosequencing of 16S rRNA gene fragments as used here is a fast and reliable method well suited for identification to the species level, even within the Lactobacillus acidophilus complex.

Keyword
Lactobacillus, vaginal fluid, 16S rRNA genes, pyrosequencing
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-13052 (URN)10.1034/j.1600-0463.2002.1101106.x (DOI)
Available from: 2008-03-19 Created: 2008-03-19 Last updated: 2016-06-30Bibliographically approved
3. An international study of the inter-observer variation between the interpretations of vaginal smear criteria of Bacterial Vaginosis
Open this publication in new window or tab >>An international study of the inter-observer variation between the interpretations of vaginal smear criteria of Bacterial Vaginosis
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2002 (English)In: APMIS, ISSN 1600-0463, Vol. 110, no 11, 811-818 p.Article in journal (Refereed) Published
Abstract [en]

An international workshop on vaginal smear-based diagnosis of bacterial vaginosis was organized where 13 investigators scoring 258 slides with smears from vaginal fluid. Interobserver reproducibility of interpretations of Nugent scores, Hay/Ison scores and wet smear scores for the diagnosis of bacterial vaginosis was shown to be high. Detailed analysis of individual scoring results however indicated that basic standards of quality control to ensure robust individual readings of slides must be adhered to.

Keyword
Bacterial vaginosis, diagnosis, criteria
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-13050 (URN)10.1034/j.1600-0463.2002.1101107.x (DOI)
Available from: 2008-03-19 Created: 2008-03-19 Last updated: 2013-09-12

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