Gardnerella vaginalis and Lactobacillus sp in liquid-based cervical samples in healthy and disturbed vaginal flora using cultivation-independent methods.
Our objective was to determine the morphotype of the adherent bacteria in liquid-based cytology (LBC) in smears with healthy and disturbed vaginal flora. And to use PCR technology on the same fixed cell sample to establish DNA patterns of the 16S RNA genes of the bacteria in the sample. Thirty samples were randomly selected from a large group of cervical cell samples suspended in a commercial coagulant fixative "(BoonFix)." PCR was used to amplify DNA of five bacterial species: Lactobacillus acidophilus, Lactobacillus crispatus, Lactobacillus jensenii, Gardnerella vaginalis, and Mycoplasma hominis. The LBC slides were then analyzed by light microscopy to estimate bacterial adhesion. DNA of lactobacilli was detected in all cell samples. Seventeen smears showed colonization with Gardnerella vaginalis (range 2.6 x 10(2)-3.0 x 10(5) bacteria/mul BoonFix sample). Two cases were identified as dysbacteriotic with high DNA values for Gardnerella vaginalis and low values for Lactobacillus crispatus. The sample with the highest concentration for Gardnerella vaginalis showed an unequivocal Gardnerella infection. This study indicates that the adherence pattern of a disturbed flora in liquid-based cervical samples can be identified unequivocally, and that these samples are suitable for quantitative PCR analysis. This cultivation independent method reveals a strong inverse relationship between Gardnerella vaginalis and Lactobacillus crispatus in dysbacteriosis and unequivocal Gardnerella infection.
Klomp
Johanna M
JM
Leiden Cytology and Pathology Laboratory, Leiden, the Netherlands.
Verbruggen
Banut-Sabine M
BS
Korporaal
Hans
H
Boon
Mathilde E
ME
de Jong
Pauline
P
Kramer
Gerco C
GC
van Haaften
Maarten
M
Heintz
A Peter M
AP
D000293:Adolescent; D000328:Adult; D001422:Bacterial Adhesion; D001431:Bacteriological Techniques; D002584:Cervix Uteri / Q000382:microbiology; D004269:DNA, Bacterial / Q000032:analysis; D004311:Double-Blind Method; D005260:Female; D006194:Gardnerella vaginalis / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007778:Lactobacillus / Q000235:genetics / Q000302:isolation & purification*; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D014621:Vagina / Q000382:microbiology*; D014626:Vaginal Smears; D016585:Vaginosis, Bacterial / Q000382:microbiology*
D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S