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An international study of the inter-observer variation between the interpretations of vaginal smear criteria of Bacterial Vaginosis
Linköping University, Department of Clinical and Experimental Medicine, Clinical Microbiology . Linköping University, Faculty of Health Sciences.
Linköping University, Department of Clinical and Experimental Medicine, Obstetrics and gynecology . Linköping University, Faculty of Health Sciences.
Dept. of Medical Microbiology, Imperial College School of Medicine at St. Mary's, London, UK.
Dept. of GU Medicine & Sexual Health, Royal Cornwall Hospital, Treliske, Truro, Great Britain.
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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.

Place, publisher, year, edition, pages
2002. Vol. 110, no 11, 811-818 p.
Keyword [en]
Bacterial vaginosis, diagnosis, criteria
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:liu:diva-13050DOI: 10.1034/j.1600-0463.2002.1101107.xOAI: oai:DiVA.org:liu-13050DiVA: diva2:17738
Available from: 2008-03-19 Created: 2008-03-19 Last updated: 2013-09-12
In thesis
1. Lactobacillus iners and the normal vaginal flora
Open this publication in new window or tab >>Lactobacillus iners and the normal vaginal flora
2008 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [fi]

The ecological niche of the vagina contains a large number of different microbes that are constantly interacting with each other and the host. Culture methods have not been sufficient in order to resolve the complexity of the normal vaginal flora. Further, the methods for delineating normal flora from not normal flora are not easily handled and are traditionally not based on culture but on microscopy of elements of the vaginal fluid. In the work presented in this thesis, an international collaboration was established that pin-pointed some of the difficulties in classifying vaginal floras, including staining, sampling, and discordance when lactobacilli are few in number, and that emphasized the importance of the size of the vision field in microscopes. As lactobacilli are prominent members of the normal vaginal flora they need to be carefully classified if further work towards more robust scoring tools is to be achieved.

Phenotypic methods have not been able to separate the closely related Lactobacillus species of the vagina. Progress in molecular biology has provided possibilities to characterize these lactobacilli, which are mainly from the Lactobacillus acidophilus group. In this work a large number of strains collected by true random sampling were subjected to RAPD-PCR, TTGE and multiplex PCR for species identification. The major species found were L. crispatus, L. gasseri and L. jensenii and the recently described L. iners. The presence of L. iners has not been detected in previous studies due to its special nutrient requirements. Development of pyrosequencing technology also made it possible to match signatures of the two variable regions V1 and V3 of the 16S rRNA gene of the vaginal lactobacilli and identify them to the species level in a high throughput manner. The study confirmed that the dominating flora in women with normal vaginal flora comprises the four species mentioned previously. Repetitive sampling during IVF-treatment with highly varying oestrogen levels demonstrates changes that possibly occur during changes in the natural life cycle. Furthermore, L. iners was found to be the first species to be established after spontaneously resolved or treated Bacterial Vaginosis.

These findings can be of help in developing new strategies for regaining and retaining the normal vaginal flora.

Place, publisher, year, edition, pages
Linköping University Electronic Press, 2008
Series
Linköping University Medical Dissertations, ISSN 0345-0082 ; 1049
Keyword
Vagina, microbes, lactobacilli, phenotypic methods, molecular biology, 16S rRNA, pyrosequencing
National Category
Microbiology in the medical area
Identifiers
urn:nbn:se:liu:diva-11334 (URN)978-91-7393-953-9 (ISBN)
Public defence
2008-04-18, Berzeliussalen, Campus US, Linköpings universitet, Linköping, 13:00 (English)
Opponent
Supervisors
Available from: 2008-03-19 Created: 2008-03-19 Last updated: 2015-11-19
2. Lactobacilli dominating the normal vaginal flora
Open this publication in new window or tab >>Lactobacilli dominating the normal vaginal flora
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:nbn:se:liu:diva-26680 (URN)11247 (Local ID)91-7373-544-2 (ISBN)11247 (Archive number)11247 (OAI)
Presentation
2003-05-15, Flemingsalen, Universitetssjukhuset, Linköping, 13:00 (Swedish)
Available from: 2009-10-08 Created: 2009-10-08 Last updated: 2013-09-12

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Forsum, UrbanJakobsson, TellCarlsson, Bodil

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