Cargando…
Cloning of 16S rRNA genes amplified from normal and disturbed vaginal microflora suggests a strong association between Atopobium vaginae, Gardnerella vaginalis and bacterial vaginosis
BACKGROUND: The pathogenesis of bacterial vaginosis remains largely elusive, although some microorganisms, including Gardnerella vaginalis, are suspected of playing a role in the etiology of this disorder. Recently culture-independent analysis of microbial ecosystems has proven its efficacy in chara...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2004
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC419343/ https://www.ncbi.nlm.nih.gov/pubmed/15102329 http://dx.doi.org/10.1186/1471-2180-4-16 |
_version_ | 1782121431892492288 |
---|---|
author | Verhelst, Rita Verstraelen, Hans Claeys, Geert Verschraegen, Gerda Delanghe, Joris Van Simaey, Leen De Ganck, Catharine Temmerman, Marleen Vaneechoutte, Mario |
author_facet | Verhelst, Rita Verstraelen, Hans Claeys, Geert Verschraegen, Gerda Delanghe, Joris Van Simaey, Leen De Ganck, Catharine Temmerman, Marleen Vaneechoutte, Mario |
author_sort | Verhelst, Rita |
collection | PubMed |
description | BACKGROUND: The pathogenesis of bacterial vaginosis remains largely elusive, although some microorganisms, including Gardnerella vaginalis, are suspected of playing a role in the etiology of this disorder. Recently culture-independent analysis of microbial ecosystems has proven its efficacy in characterizing the diversity of bacterial populations. Here, we report on the results obtained by combining culture and PCR-based methods to characterize the normal and disturbed vaginal microflora. RESULTS: A total of 150 vaginal swab samples from healthy women (115 pregnant and 35 non-pregnant) were categorized on the basis of Gram stain of direct smear as grade I (n = 112), grade II (n = 26), grade III (n = 9) or grade IV (n = 3). The composition of the vaginal microbial community of eight of these vaginal swabs (three grade I, two grade II and three grade III), all from non-pregnant women, were studied by culture and by cloning of the 16S rRNA genes obtained after direct amplification. Forty-six cultured isolates were identified by tDNA-PCR, 854 cloned 16S rRNA gene fragments were analysed of which 156 by sequencing, yielding a total of 38 species, including 9 presumptively novel species with at least five species that have not been isolated previously from vaginal samples. Interestingly, cloning revealed that Atopobium vaginae was abundant in four out of the five non-grade I specimens. Finally, species specific PCR for A. vaginae and Gardnerella vaginalis pointed to a statistically significant co-occurrence of both species in the bacterial vaginosis samples. CONCLUSIONS: Although historically the literature regarding bacterial vaginosis has largely focused on G. vaginalis in particular, several findings of this study – like the abundance of A. vaginae in disturbed vaginal microflora and the presence of several novel species – indicate that much is to be learned about the composition of the vaginal microflora and its relation to the etiology of BV. |
format | Text |
id | pubmed-419343 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2004 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-4193432004-05-28 Cloning of 16S rRNA genes amplified from normal and disturbed vaginal microflora suggests a strong association between Atopobium vaginae, Gardnerella vaginalis and bacterial vaginosis Verhelst, Rita Verstraelen, Hans Claeys, Geert Verschraegen, Gerda Delanghe, Joris Van Simaey, Leen De Ganck, Catharine Temmerman, Marleen Vaneechoutte, Mario BMC Microbiol Research Article BACKGROUND: The pathogenesis of bacterial vaginosis remains largely elusive, although some microorganisms, including Gardnerella vaginalis, are suspected of playing a role in the etiology of this disorder. Recently culture-independent analysis of microbial ecosystems has proven its efficacy in characterizing the diversity of bacterial populations. Here, we report on the results obtained by combining culture and PCR-based methods to characterize the normal and disturbed vaginal microflora. RESULTS: A total of 150 vaginal swab samples from healthy women (115 pregnant and 35 non-pregnant) were categorized on the basis of Gram stain of direct smear as grade I (n = 112), grade II (n = 26), grade III (n = 9) or grade IV (n = 3). The composition of the vaginal microbial community of eight of these vaginal swabs (three grade I, two grade II and three grade III), all from non-pregnant women, were studied by culture and by cloning of the 16S rRNA genes obtained after direct amplification. Forty-six cultured isolates were identified by tDNA-PCR, 854 cloned 16S rRNA gene fragments were analysed of which 156 by sequencing, yielding a total of 38 species, including 9 presumptively novel species with at least five species that have not been isolated previously from vaginal samples. Interestingly, cloning revealed that Atopobium vaginae was abundant in four out of the five non-grade I specimens. Finally, species specific PCR for A. vaginae and Gardnerella vaginalis pointed to a statistically significant co-occurrence of both species in the bacterial vaginosis samples. CONCLUSIONS: Although historically the literature regarding bacterial vaginosis has largely focused on G. vaginalis in particular, several findings of this study – like the abundance of A. vaginae in disturbed vaginal microflora and the presence of several novel species – indicate that much is to be learned about the composition of the vaginal microflora and its relation to the etiology of BV. BioMed Central 2004-04-21 /pmc/articles/PMC419343/ /pubmed/15102329 http://dx.doi.org/10.1186/1471-2180-4-16 Text en Copyright © 2004 Verhelst et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL. |
spellingShingle | Research Article Verhelst, Rita Verstraelen, Hans Claeys, Geert Verschraegen, Gerda Delanghe, Joris Van Simaey, Leen De Ganck, Catharine Temmerman, Marleen Vaneechoutte, Mario Cloning of 16S rRNA genes amplified from normal and disturbed vaginal microflora suggests a strong association between Atopobium vaginae, Gardnerella vaginalis and bacterial vaginosis |
title | Cloning of 16S rRNA genes amplified from normal and disturbed vaginal microflora suggests a strong association between Atopobium vaginae, Gardnerella vaginalis and bacterial vaginosis |
title_full | Cloning of 16S rRNA genes amplified from normal and disturbed vaginal microflora suggests a strong association between Atopobium vaginae, Gardnerella vaginalis and bacterial vaginosis |
title_fullStr | Cloning of 16S rRNA genes amplified from normal and disturbed vaginal microflora suggests a strong association between Atopobium vaginae, Gardnerella vaginalis and bacterial vaginosis |
title_full_unstemmed | Cloning of 16S rRNA genes amplified from normal and disturbed vaginal microflora suggests a strong association between Atopobium vaginae, Gardnerella vaginalis and bacterial vaginosis |
title_short | Cloning of 16S rRNA genes amplified from normal and disturbed vaginal microflora suggests a strong association between Atopobium vaginae, Gardnerella vaginalis and bacterial vaginosis |
title_sort | cloning of 16s rrna genes amplified from normal and disturbed vaginal microflora suggests a strong association between atopobium vaginae, gardnerella vaginalis and bacterial vaginosis |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC419343/ https://www.ncbi.nlm.nih.gov/pubmed/15102329 http://dx.doi.org/10.1186/1471-2180-4-16 |
work_keys_str_mv | AT verhelstrita cloningof16srrnagenesamplifiedfromnormalanddisturbedvaginalmicroflorasuggestsastrongassociationbetweenatopobiumvaginaegardnerellavaginalisandbacterialvaginosis AT verstraelenhans cloningof16srrnagenesamplifiedfromnormalanddisturbedvaginalmicroflorasuggestsastrongassociationbetweenatopobiumvaginaegardnerellavaginalisandbacterialvaginosis AT claeysgeert cloningof16srrnagenesamplifiedfromnormalanddisturbedvaginalmicroflorasuggestsastrongassociationbetweenatopobiumvaginaegardnerellavaginalisandbacterialvaginosis AT verschraegengerda cloningof16srrnagenesamplifiedfromnormalanddisturbedvaginalmicroflorasuggestsastrongassociationbetweenatopobiumvaginaegardnerellavaginalisandbacterialvaginosis AT delanghejoris cloningof16srrnagenesamplifiedfromnormalanddisturbedvaginalmicroflorasuggestsastrongassociationbetweenatopobiumvaginaegardnerellavaginalisandbacterialvaginosis AT vansimaeyleen cloningof16srrnagenesamplifiedfromnormalanddisturbedvaginalmicroflorasuggestsastrongassociationbetweenatopobiumvaginaegardnerellavaginalisandbacterialvaginosis AT deganckcatharine cloningof16srrnagenesamplifiedfromnormalanddisturbedvaginalmicroflorasuggestsastrongassociationbetweenatopobiumvaginaegardnerellavaginalisandbacterialvaginosis AT temmermanmarleen cloningof16srrnagenesamplifiedfromnormalanddisturbedvaginalmicroflorasuggestsastrongassociationbetweenatopobiumvaginaegardnerellavaginalisandbacterialvaginosis AT vaneechouttemario cloningof16srrnagenesamplifiedfromnormalanddisturbedvaginalmicroflorasuggestsastrongassociationbetweenatopobiumvaginaegardnerellavaginalisandbacterialvaginosis |