title: Unravelling the Bacterial Vaginosis-Associated Biofilm: A Multiplex Gardnerella vaginalis and Atopobium vaginae Fluorescence In Situ Hybridization Assay Using Peptide Nucleic Acid Probes.
abstract: Bacterial vaginosis (BV), a condition defined by increased vaginal discharge without significant inflammation, is characterized by a change in the bacterial composition of the vagina. Lactobacillus spp., associated with a healthy vaginal microbiome, are outnumbered by BV-associated organisms. These bacteria could form a polymicrobial biofilm which allows them to persist in spite of antibiotic treatment. In this study, we examined the presence of Gardnerella vaginalis and Atopobium vaginae in vaginal biofilms using Peptide Nucleic Acid (PNA) probes targeting these bacteria. For this purpose, we developed three new PNA probes for A. vaginae. The most specific A. vaginae probe, AtoITM1, was selected and then used in an assay with two existing probes, Gard162 and BacUni-1, to evaluate multiplex FISH on clinical samples. Using quantitative polymerase chain reaction (qPCR) as the gold standard, we demonstrated a sensitivity of 66.7% (95% confidence interval: 54.5% - 77.1%) and a specificity of 89.4% (95% confidence interval: 76.1% - 96%) of the new AtoITM1 probe. FISH enabled us to show the presence of a polymicrobial biofilm in bacterial vaginosis, in which Atopobium vaginae is part of a Gardnerella vaginalis-dominated biofilm. We showed that the presence of this biofilm is associated with high bacterial loads of A. vaginae and G. vaginalis.
authors: Unit of Epidemiology and Control of HIV/STD, Department of Public Health, Institute of Tropical Medicine, Antwerp, Belgium; Laboratory Bacteriology Research, Faculty of Medicine & Health Sciences, University of Ghent, Ghent, Belgium; STI Reference Laboratory, Department of Clinical Sciences, Institute of Tropical Medicine, Antwerp, Belgium.
authors: Liselotte
authors: L
authors: Hardy
authors: Unit of Epidemiology and Control of HIV/STD, Department of Public Health, Institute of Tropical Medicine, Antwerp, Belgium.
authors: Vicky
authors: V
authors: Jespers
authors: STI Reference Laboratory, Department of Clinical Sciences, Institute of Tropical Medicine, Antwerp, Belgium; Plantijn Hogeschool, Antwerp, Belgium.
authors: Nassira
authors: N
authors: Dahchour
authors: Rinda Ubuzima, Kigali, Rwanda.
authors: Lambert
authors: L
authors: Mwambarangwe
authors: Rinda Ubuzima, Kigali, Rwanda.
authors: Viateur
authors: V
authors: Musengamana
authors: Laboratory Bacteriology Research, Faculty of Medicine & Health Sciences, University of Ghent, Ghent, Belgium.
authors: Mario
authors: M
authors: Vaneechoutte
authors: STI Reference Laboratory, Department of Clinical Sciences, Institute of Tropical Medicine, Antwerp, Belgium.
authors: Tania
authors: T
authors: Crucitti
mesh_terms: D039903:Actinobacteria / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D018441:Biofilms; D005260:Female; D006194:Gardnerella vaginalis / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D015341:Nucleic Acid Probes / Q000235:genetics; D020135:Peptide Nucleic Acids / Q000235:genetics; D014621:Vagina / Q000382:microbiology; D016585:Vaginosis, Bacterial / Q000235:genetics / Q000382:microbiology* / Q000473:pathology
chemical_list: D015341:Nucleic Acid Probes; D020135:Peptide Nucleic Acids