Donald School Journal of Ultrasound in Obstetrics and Gynecology

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VOLUME 16 , ISSUE 2 ( April-June, 2022 ) > List of Articles

Original Scientific Article

Molecular Analysis of Vaginal Microbiome in Women of Reproductive Age

Kornelija Trajkova, Milena Petrovska, Aleksandar Dimovski, Biljana Curcic, Katerina Popovska Jankovic, Gordana Adamova

Keywords : Bacterial vaginosis, Lactobacillus, Molecular analysis, Vaginal microbiome

Citation Information : Trajkova K, Petrovska M, Dimovski A, Curcic B, Jankovic KP, Adamova G. Molecular Analysis of Vaginal Microbiome in Women of Reproductive Age. Donald School J Ultrasound Obstet Gynecol 2022; 16 (2):107-117.

DOI: 10.5005/jp-journals-10009-1932

License: CC BY-NC 4.0

Published Online: 05-07-2022

Copyright Statement:  Copyright © 2022; The Author(s).


Aim: Molecular analysis of the vaginal microbiome (VM) in terms of types of Lactobacilli and their role. Material and methods: Ninety-three women of reproductive age were included in a prospective cross-sectional study. A vaginal examination and collection of two vaginal swabs, first for Nugent score and second for molecular identification of Lactobacillus spp. and other bacteria were performed. Identification was done by polymerase chain reaction (PCR) amplification of 16S rRNK genes and sequencing for Lactobacillus types. Results: Twenty-two species of Lactobacillus have been identified, 13 types in all 51 participants of the normal flora (NF) group, 19 types in 95.6% intermediate group (IM) (n = 23), and only seven in 68.4% of women in bacterial vaginosis (BV) (n = 19) group. Three types were the most common: Lactobacillus iners, almost equally present in all three groups, predominantly as a single type and in combination with other pathogenic bacteria (Gardnerella vaginalis and two types of Mycoplasmataceae), and Lactobacillus crispatus and Lactobacillus casei, both significantly associated with NF. L. crispatus was more abundant in the secretory vs proliferative phase of menstrual cycle. Conclusion: Colonization with any Lactobacillus and especially L. crispatus and L. casei, is generally significantly associated with NF. L. iners is associated with dysbiosis. The molecular analysis of the VM may significantly participate in developing strategies for prevention and treatment of genital infections.

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  1. Peterson J, Garges S, Giovanni M, et al. The NIH Human Microbiome Project. Genome Res 2009;19(12):2317–2323. DOI: 10.1101/gr.096651.109
  2. Lederberg J, McCray AT. ‘Ome sweet ‘omics—a genealogical treasury of words. Genealogical Treasury of Words. Scientist 2001;15(7):8. Available at: (accessed 18 June, 2022)
  3. Reid G. Probiotic agents to protect the urogenital tract against infection. Am J Clin Nutr 2001;73(2 Suppl):437S–443S. DOI: 10.1093/ajcn/73.2.437s
  4. Hillier SL, Krohn MA, Klebanoff SJ, et al. The relationship of hydrogen peroxide-producing lactobacilli to bacterial vaginosis and genital microflora in pregnant women. Obstet Gynecol 1992;79(3):369–373. DOI: 10.1097/00006250-199203000-00008
  5. Antonio MA, Rabe LK, Hillier SL. Colonization of the rectum by Lactobacillus species and decreased risk of bacterial vaginosis. J Infect Dis 2005;192(3):394–398. DOI: 10.1086/430926
  6. Yeruva T, Rajkumar H, Donugama V. Vaginal lactobacilli profile in pregnant women with normal & abnormal vaginal flora. Indian J Med Res 2017;146(4):534–540. DOI: 10.4103/ijmr.IJMR_774_16
  7. Tabatabaei N, Eren AM, Barreiro LB, et al. Vaginal microbiome in early pregnancy and subsequent risk of spontaneous preterm birth: a case-control study. BJOG 2019;126(3):349–358. DOI: 10.1111/1471-0528.15299
  8. Onderdonk AB, Delaney ML, Fichorova RN. The human microbiome during bacterial vaginosis. Clin Microbiol Rev 2016;29(2):223–238. DOI: 10.1128/CMR.00075-15
  9. Saling E, Schreiber M, al-Taie T. A simple, efficient and inexpensive program for preventing prematurity. J Perinat Med 2001;29(3):199–211. DOI: 10.1515/JPM.2001.029
  10. Saling E, Fuhr N, Placht A, et al. A new efficient strategy for prevention of prematurity. In: Kurjak A, Latin V, Rippmann E, editors. Advances on the Pathophysiology of Pregnancy. Rome: CIC Edizioni Internationali; 1995. p. 228–234.
  11. Saling E. Problems in prevention of preterm birth—regrettable contradictions. A special comment from the Founder of the Journal of Perinatal Medicine. J Perinat Med 2011;39(3):223–225. DOI: 10.1515/jpm.2011.033
  12. Nugent RP, Krohn MA, Hillier SL. Reliability of diagnosing bacterial vaginosis is improved by a standardized method of gram stain interpretation. J Clin Microbiol 1991;29(2):297–301. DOI: 10.1128/jcm.29.2.297-301.1991
  13. Efremov GD, Dimovski AJ, Plaseska-Karanfilska D, et al. Laboratory Manual. 3rd ed. RCGEB, MASA; 1999.
  14. Tamrakar R, Yamada T, Furuta I, et al. Association between Lactobacillus species and bacterial vaginosis-related bacteria, and bacterial vaginosis scores in pregnant Japanese women. BMC Infect Dis 2007;7:128. DOI: 10.1186/1471-2334-7-128
  15. Peerayeh NS, Sattari M. Detection of Ureaplasma urealyticum and Mycoplasma hominis in endocervical specimens from infertile women by polymerase chain reaction. Middle East Fertil Soc J 2006;11(2):104–108. Available at: (accessed 18 June, 2022)
  16. Gergova RT, Strateva TV, Mitov IG. Gardnerella vaginalis-associated bacterial vaginosis in Bulgarian women. Braz J Infect Dis 2013;17(3):313–318. DOI: 10.1016/j.bji d.2012.10.026
  17. Zhou X, Hansmann MA, Davis CC, et al. The vaginal bacterial communities of Japanese women resemble those of women in other racial groups. FEMS Immunol Med Microbiol 2010;58(2):169–181. DOI: 10.1111/j.1574-695X.2009.00618.x
  18. Damelin LH, Paximadis M, Mavri-Damelin D, et al. Identification of predominant culturable vaginal Lactobacillus species and associated bacteriophages from women with and without vaginal discharge syndrome in South Africa. J Med Microbiol 2011;60(Pt 2):180–183. DOI: 10.1099/jmm.0.024463-0
  19. Martínez-Peña MD, Castro-Escarpulli G, Aguilera-Arreola MG. Lactobacillus species isolated from vaginal secretions of healthy and bacterial vaginosis-intermediate Mexican women: a prospective study. BMC Infect Dis 2013;13:189. DOI: 10.1186/1471-2334-13-189
  20. Pendharkar S, Magopane T, Larsson PG, et al. Identification and characterization of vaginal lactobacilli from South African women. BMC Infect Dis 2013;13:43. DOI: 10.1186/1471-2334-13-43
  21. Smith SB, Ravel J. The vaginal microbiota, host defence and reproductive physiology. J Physiol 2017;595(2):451–463. DOI: 10.1113/JP271694
  22. López-Filloy M, Cortez FJ, Gheit T, et al. Altered vaginal microbiota composition correlates with human papillomavirus and mucosal immune responses in women with symptomatic cervical ectopy. Front Cell Infect Microbiol 2022;12:884272. DOI: 10.3389/fcimb.2022.884272
  23. Song YL, Kato N, Matsumiya Y, et al. Identification of and hydrogen peroxide production by fecal and vaginal lactobacilli isolated from Japanese women and newborn infants. J Clin Microbiol 1999;37(9):3062–3064. DOI: 10.1128/JCM.37.9.3062-3064.1999
  24. Vallor AC, Antonio MA, Hawes SE, et al. Factors associated with acquisition of, or persistent colonization by, vaginal lactobacilli: role of hydrogen peroxide production. J Infect Dis 2001;184(11):1431–1436. DOI: 10.1086/324445
  25. Pavlova SI, Kilic AO, Kilic SS, et al. Genetic diversity of vaginal lactobacilli from women in different countries based on 16S rRNA gene sequences. J Appl Microbiol 2002;92(3):451–459. DOI: 10.1046/j.1365-2672.2002.01547.x
  26. Vásquez A, Jakobsson T, Ahrné S, et al. Vaginal lactobacillus flora of healthy Swedish women. J Clin Microbiol 2002;40(8): 2746–2749. DOI: 10.1128/JCM.40.8.2746-2749.2002
  27. Verstraelen H, Verhelst R, Claeys G, et al. Longitudinal analysis of the vaginal microflora in pregnancy suggests that L. crispatus promotes the stability of the normal vaginal microflora and that L. gasseri and/or L. iners are more conducive to the occurrence of abnormal vaginal microflora. BMC Microbiol 2009;9:116. DOI: 10.1186/1471-2180-9-116
  28. Jakobsson T, Forsum U. Lactobacillus iners: a marker of changes in the vaginal flora? J Clin Microbiol 2007;45(9):3145. DOI: 10.1128/JCM.00558-07
  29. Srinivasan S, Liu C, Mitchell CM, et al. Temporal variability of human vaginal bacteria and relationship with bacterial vaginosis. PLoS One 2010;5(4):e10197. DOI: 10.1371/journal.pone.0010197
  30. Petrova MI, Lievens E, Malik S, et al. Lactobacillus species as biomarkers and agents that can promote various aspects of vaginal health. Front Physiol 2015;6:81. DOI: 10.3389/fphys.2015.00081
  31. Borgdorff H, Armstrong SD, Tytgat HL, et al. Unique insights in the cervicovaginal Lactobacillus iners and L. crispatus proteomes and their associations with microbiota dysbiosis. PLoS One 2016;11(3):e0150767. DOI: 10.1371/journal.pone.0150767
  32. Fosch SE, Ficoseco CA, Marchesi A, et al. Contraception: influence on vaginal microbiota and identification of vaginal lactobacilli using MALDI-TOF MS and 16S rDNA sequencing. Open Microbiol J 2018;12:218–229. DOI: 10.2174/1874285801812010218
  33. Crucitti T, Hardy L, van de Wijgert J, et al. Contraceptive rings promote vaginal lactobacilli in a high bacterial vaginosis prevalence population: a randomised, open-label longitudinal study in Rwandan women. PLoS One 2018;13(7):e0201003. DOI: 10.13039/501100001713
  34. De Seta F, Restaino S, De Santo D, et al. Effects of hormonal contraception on vaginal flora. Contraception 2012;86(5): 526–529. DOI: 10.1016/j.contraception.2012.02.012
  35. Jie Z, Chen C, Hao L, et al. Life history recorded in the vagino-cervical microbiome along with multi-omics. Genomics Proteomics Bioinformatics 2021;9:S1672-0229(21)00099-1 DOI: 10.1016/j.gpb.2021.01.005. Epub ahead of print. PMID: 34118463.
  36. Gajer P, Brotman RM, Bai G, et al. Temporal dynamics of the human vaginal microbiota. Sci Transl Med 2012;4(132):132ra52 DOI: 10.1126/scitranslmed.3003605
  37. Nasioudis D, Forney LJ, Schneider GM, et al. Influence of pregnancy history on the vaginal microbiome of pregnant women in their first trimester. Sci Rep 2017;7(1):10201. DOI: 10.1038/s41598-017-09857-z
  38. Martin DH. The microbiota of the vagina and its influence on women's health and disease. Am J Med Sci 2012;343(1):2–9. DOI: 10.1097/MAJ.0b013e31823ea228
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