Comparison of methods for detecting biofilm syndrome in bacterial vaginosis

DOI

https://dx.doi.org/10.18565/aig.2025.117

Shalepo K.V., Spasibova E.V., Krysanova A.A., Khusnutdinova T.A., Budilovskaya O.V., Storozheva K.V., Tapilskaya N.I., Savicheva A.M., Kogan I.Yu.
D.O. Ott Research Institute for Obstetrics, Gynecology and Reproductology, St. Petersburg, Russia

In bacterial vaginosis, Gardnerella vaginalis and Fannyhessea vaginae form highly structured, dense biofilm consortia. Key cells serve as markers of biofilm organization within bacterial communities.
Objective: To compare microbiological methods for detecting biofilm syndrome in bacterial vaginosis.
Materials and methods: Twenty-eight women with complaints of vaginal discharge were examined. Vaginal discharge was used as the clinical material. Microscopy of preparations stained with Gram and methylene blue, as well as fluorescent in situ hybridization (FISH) or the RiGinaM method, were used. The Femoflor test was used for molecular analysis.
Results: The diagnosis of bacterial vaginosis in the Nugent study was established in 89.3% (25/28) of women. The microscopic evaluation of vaginal microbiocenosis using aniline dyes and the RiGinaM method yielded identical results (L:E ratio less than 4:1, with clue cells detected). Clue cells were not identified by any of the methods in only three cases. Among the 25 patients diagnosed with bacterial vaginosis by the Femoflor test, G. vaginalis was detected in all clinical samples, with an average logarithmic value of 7.55, whereas F. vaginae was detected in 22 cases. In three instances of physiological vaginal microbiocenosis, the total bacterial mass was predominantly represented by lactobacilli. G. vaginalis was found among these women, but their total bacterial mass was low (3.6, 4.1, and 6.8 lg), and F. vaginae was detected in only one case (2.1 lg).
Conclusion: Clue cells, as markers of biofilm bacterial vaginosis, can be detected using Gram staining, methylene blue staining, and the RiGinaM method. Microscopic methods using aniline dyes do not differentiate the types of bacteria present in the clue cells, whereas the RiGinaM method requires specific probes to identify microorganisms. The multiplex PCR method in real time can be utilized for the diagnosis of bacterial vaginosis in conjunction with microscopy methods.

Authors' contributions: Shalepo K.V. – methodology, design, and coordination of the study, drafting and editing of the manuscript; Spasibova E.V. – methodology, design, and coordination of the study; Storozheva K.V. – conducting the study, participation drafting and editing of the manuscript; Khusnutdinova T.A., Budilovskaya O.V., Krysanova A.A. – participation in drafting and editing of the manuscript; Tapilskaya N.I., Savicheva A.M. – conception and coordination of the study, editing of the manuscript, approval of the final version of the manuscript; Kogan I.Yu. – conception of the study. All authors contributed significantly to the research and analysis and drafting of the manuscript, read and approved the final version prior to submission.
Conflicts of interest: The authors have no conflicts of interest to declare.
Funding: The study was conducted within the framework of the Federal Research Institute's scientific topic, "Universal and Specific Mechanisms of Implementation and Disorders of Reproductive Function in the Family", reg. No. 1021062812133-0-3.2.2.
Ethical Approval: The study was reviewed and approved by the Research Ethics Committee of D.O. Ott Research Institute for OG&R.
Patient Consent for Publication: All patients provided informed consent for the publication of their data.
Authors' data sharing statement: The data supporting the findings of this study are available upon request from the corresponding author after approval from the principal investigator.
For citation: Shalepo K.V., Spasibova E.V., Krysanova A.A., Khusnutdinova T.A., Budilovskaya O.V.,
Storozheva K.V., Tapilskaya N.I., Savicheva A.M., Kogan I.Yu. Comparison of
methods for detecting biofilm syndrome in bacterial vaginosis.
Akusherstvo i Ginekologiya/Obstetrics and Gynecology. 2025; (7): 121-129 (in Russian)
https://dx.doi.org/10.18565/aig.2025.117

bacterial vaginosis

biofilm syndrome

biofilms

clue cells

microscopy

RiGinaM

Full text (in Russian)

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Received 30.04.2025

Accepted 18.06.2025

Kira V. Shalepo, PhD, Senior Researcher at the Experimental Microbiology Group, D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology,
3 Mendeleyevskaya Line, 199034, St. Petersburg, Russia, +7(911)2474151, 2474151@mail.ru, https://orcid.org/ 0000-0002-3002-3874
Elena V. Spasibova, Bacteriologist at the Laboratory of Clinical Microbiology, D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology, 3 Mendeleyevskaya Line, 199034, St. Petersburg, Russia, elena.graciosae@gmail.com, https://orcid.org/0009-0002-6070-4651
Anna A. Krysanova, PhD, Senior Researcher at the Experimental Microbiology Group, D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology,
3 Mendeleyevskaya Line, 199034, St. Petersburg, Russia, krusanova.anna@mail.ru, https://orcid.org/ 0000-0003-4798-1881
Olga V. Budilovskaya, PhD, Senior Researcher at the Experimental Microbiology Group, D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology,
3 Mendeleyevskaya Line, 199034, St. Petersburg, Russia, o.budilovskaya@gmail.com, https://orcid.org/0000-0001-7673-6274
Tatiana A. Khusnutdinova, PhD, Senior Researcher at the Experimental Microbiology Group, D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology,
3 Mendeleyevskaya Line, 199034, St. Petersburg, Russia, husnutdinovat@yandex.ru, https://orcid.org/0000-0002-2742-2655
Kseniia V. Storozheva, PhD Student, D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology, 3 Mendeleyevskaya Line,
199034, St. Petersburg, Russia, kvstorozheva@mail.ru, https://orcid.org/0009-0005-8954-0234
Natalya I. Tapilskaya, Dr. Med. Sci., Professor, Leading Researcher at the Reproduction Department, D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology, 3 Mendeleyevskaya Line, 199034, St. Petersburg, Russia, tapnatalia@mail.ru, https://orcid.org/ 0000-0001-5309-0087
Alevtina M. Savicheva, Dr. Med. Sci., Professor, Head of the Department of Medical Microbiology, D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology, 3 Mendeleyevskaya Line, 199034, St. Petersburg, Russia, savitcheva@mail.ru, https://orcid.org/ 0000-0003-3870-5930
Igor Yu. Kogan, Corresponding Member of the RAS, Dr. Med. Sci., Professor, Director, D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology,
3 Mendeleyevskaya Line, 199034, St. Petersburg, Russia, ovr@ott.ru, https://orcid.org/0000-0002-7351-6900
Corresponding author: Kira V. Shalepo, 2474151@mail.ru

Comparison of methods for detecting biofilm syndrome in bacterial vaginosis

Shalepo K.V., Spasibova E.V., Krysanova A.A., Khusnutdinova T.A., Budilovskaya O.V., Storozheva K.V., Tapilskaya N.I., Savicheva A.M., Kogan I.Yu.

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