Phenotypic and molecular identification of lactobacilli isolated from vaginal secretions.
OBJECTIVE
Twenty lactobacilli isolated from vaginal samples of healthy volunteers were characterized according to polyphasic taxonomy. A broad spectrum of activity and protective properties of these vaginal isolates has been reported.


METHODS
Phenotypic and genotypic methods such as random amplified polymorphic DNA (RAPD), species-specific and BOX polymerase chain reaction (PCR), amplified ribosomal DNA restriction analysis (ARDRA), and 16S rRNA sequence analyses were applied for Lactobacillus spp. identification.


RESULTS
On the basis of carbohydrate utilization profiles using API 50 CHL kits (fermentation pattern), the strains were divided into 7 groups. RAPD- and BOX-PCR analyses revealed heterogeneity within the established phenotypic groups and discriminated successfully between the vaginal strains. The combination of species-specific PCR, ARDRA, and 16S rRNA sequence analyses allowed the species identification of 7 strains as Lactobacillus fermentum, 2 as Lactobacillus gasseri, 1 as Lactobacillus brevis, and 1 as Lactobacillus salivarius. These results were not in agreement with data obtained by classical and molecular methods for some clinical isolates.


CONCLUSIONS
This is the first polyphasic taxonomy study on vaginal lactobacilli from Bulgarian women of reproductive age. These data add to the limited information existing in Bulgaria on the true identity of Lactobacillus spp. in the vaginal microbiota.
Dimitonova
Silviya Petrova
SP
Department of Microbial Genetics, Institute of Microbiology, Bulgarian Academy of Sciences, Sofia, Bulgaria.
Bakalov
Boris Vladimirov
BV
Aleksandrova-Georgieva
Ralitsa Nedkova
RN
Danova
Svetla Trifonova
ST
D016000:Cluster Analysis; D004269:DNA, Bacterial / Q000032:analysis; D005260:Female; D006801:Humans; D007778:Lactobacillus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D019105:Random Amplified Polymorphic DNA Technique; D014621:Vagina / Q000382:microbiology*
D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S