Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Nov 18:11:779376.
doi: 10.3389/fcimb.2021.779376. eCollection 2021.

A New PNA-FISH Probe Targeting Fannyhessea vaginae

Lúcia G V Sousa  1 , Joana Castro  1 , Angela França  1 , Carina Almeida  1   2 , Christina A Muzny  3 , Nuno Cerca  1
Affiliations

A New PNA-FISH Probe Targeting Fannyhessea vaginae

Lúcia G V Sousa et al. Front Cell Infect Microbiol. .

Abstract

Bacterial vaginosis (BV) is the most common vaginal infection in women of reproductive age and has been associated with serious health complications, mainly in pregnant women. It is characterized by a decrease in the number of Lactobacillus species in the healthy vaginal microbiota and an overgrowth of strict and facultative anaerobic bacteria that develop a polymicrobial biofilm. Despite over 60 years of research investigating BV, its etiology is not fully understood. Gardnerella spp. is a crucial microorganism that contributes to the formation of the biofilm and the development of BV, but the role of other BV-associated bacteria is not clear. Nevertheless, Fannyhessea vaginae (previously known as Atopobium vaginae) is a highly specific species for BV, and co-colonization with Gardnerella is thought to be a very specific diagnostic marker. The diagnosis of BV still presents some limitations, since currently used methods often fail to accurately detect BV. This work aims to develop a novel peptide nucleic acid (PNA) probe targeting F. vaginae. This probe was further validated in a multiplex assay, which included a Gardnerella-specific PNA probe, as a possible method for diagnosis of BV, and was compared with quantification by qPCR. The new PNA probe showed excellent sensitivity and specificity and could discriminate F. vaginae-Gardnerella biofilms, confirming the potential to be used for the detection of BV-associated pathogens.

Keywords: Fannyhessea vaginae; Gardnerella vaginalis; bacterial vaginosis; fluorescence in situ hybridization (FISH); peptide nucleic acid (PNA).

PubMed Disclaimer

Conflict of interest statement

CM has received research grant support from Lupin Pharmaceuticals, is a consultant for Lupin Pharmaceuticals and BioFire Diagnostics, and has received honoraria from Elsevier, Abbott Molecular, Cepheid, Becton Dickinson, Roche Diagnostics, and Lupin. She is currently on the scientific advisory board for Roche and PhagoMed. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Examples of fluorescence microscopy results using the novel PNA probe with F. vaginae strains and other BV-related species. F. vaginae ATCC BAA-55 (good hybridization), F. vaginae PB2003/189-T1-4 (poor hybridization), G. vaginalis ATCC 14018, P. bivia ATCC 29303, M. curtisii ATCC 35241, L. iners ATCC 55195, P. anaerobius ATCC 27337 and L. crispatus EX533959VCO6 (absence of hybridization). For each strain/species an image of DAPI staining (DAPI filter) and the correspondent signal of the probe (FITC filter) is shown. The images were acquired with a magnification of 400×.
Figure 2
Figure 2
Confocal Laser Scanning Microscopy images of single- and dual-species biofilms hybridization with Gard162 and FvagPNA651 probes. Gard162 probe detected G. vaginalis 24 h single-species biofilm and F. vaginae probe detected F. vaginae 24 h single-species biofilm. The probes were able to detect G. vaginalis (red) and F. vaginae (green) on 24 h dual-species biofilm. The images were acquired using 40× objective.
See this image and copyright information in PMC

References

    1. Africa C. W. J. (2013). Efficacy of Methods Used for the Diagnosis of Bacterial Vaginosis. Expert Opin. Med. Diagn. 7, 189–200. doi: 10.1517/17530059.2013.753876 - DOI - PubMed
    1. Almeida C., Azevedo N. F., Bento J. C., Cerca N., Ramos H., Vieira M. J., et al. . (2013). Rapid Detection of Urinary Tract Infections Caused by Proteus Spp. Using PNA-FISH. Eur. J. Clin. Microbiol. Infect. Dis. 32, 781–786. doi: 10.1007/s10096-012-1808-2 - DOI - PubMed
    1. Almeida C., Azevedo N. F., Santos S., Keevil C. W., Vieira M. J. (2011). Discriminating Multi-Species Populations in Biofilms With Peptide Nucleic Acid Fluorescence in Situ Hybridization (PNA FISH). PloS One 6: e14786. doi: 10.1371/journal.pone.0014786 - DOI - PMC - PubMed
    1. Amsel R., Totten P. A., Spiegel C. A., Chen K. C. S., Eschenbach D., Holmes K. K. (1983). Nonspecific Vaginitis. Am. J. Med. 74, 14–22. doi: 10.1016/0002-9343(83)91112-9 - DOI - PubMed
    1. Bradshaw C. S., Morton A. N., Hocking J., Garland S. M., Morris M. B., Moss L. M., et al. . (2006. a). High Recurrence Rates of Bacterial Vaginosis Over the Course of 12 Months After Oral Metronidazole Therapy and Factors Associated With Recurrence. J. Infect. Dis. 193, 1478–1486. doi: 10.1086/503780 - DOI - PubMed

Publication types

Supplementary concepts

LinkOut - more resources