{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T17:14:28Z","timestamp":1772558068477,"version":"3.50.1"},"reference-count":34,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T00:00:00Z","timestamp":1772496000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Background\/Objectives: The cervicovaginal microbiome has emerged as a critical determinant of cervical health. In this study, we aimed to characterize the cervicovaginal microbiome across a spectrum of cervical health states and to identify community-level features that distinguish invasive disease from precursor states. Methods: We analyzed cervicovaginal samples of 86 patients with normal epithelium, low-grade (LSIL) and high-grade (HSIL) intraepithelial lesions, and cervical carcinoma (CCU) and available HPV genotyping. Vaginal samples were subjected to full-length 16S rRNA gene sequencing and genus-level taxonomic profiles were generated using ONT-supported workflows. Microbiome diversity and composition were assessed using Aitchison-based beta-diversity, non-parametric testing, and PERMANOVA. Differential abundance was evaluated using ANCOM-BC2 with false discovery rate correction. Disease-associated community shifts were quantified using log-ratio indices and co-occurrence network analysis. Results: Microbial diversity increased with disease severity, with cervical cancer showing the highest alpha diversity and distinct community composition. Normal samples were uniformly dominated by Lactobacillus, whereas LSIL and HSIL exhibited transitional communities with partial loss of lactobacillar dominance and increasing representation of anaerobic taxa. Cervical cancer was associated with depletion of Lactobacillus and expansion of anaerobic consortia. A Lactobacillus-to-anaerobe log-ratio declined monotonically with disease severity and robustly discriminated invasive cancer from precursor states. Microbial co-occurrence networks became progressively more structured with disease severity, transitioning to dense anaerobic networks in cervical cancer. Conclusions: Cervicovaginal microbiome signatures reflect cervical disease stage and may complement existing screening and risk stratification strategies.<\/jats:p>","DOI":"10.3390\/diagnostics16050753","type":"journal-article","created":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T15:22:21Z","timestamp":1772551341000},"page":"753","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Cervicovaginal Microbiome Signatures Across Cervical Disease States: A Prospective Cross-Sectional Analysis"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2494-1801","authenticated-orcid":false,"given":"Alexandru","family":"Hamod","sequence":"first","affiliation":[{"name":"Grigore T. Popa University of Medicine and Pharmacy, 700115 Iasi, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Oancea","family":"Mihaela","sequence":"additional","affiliation":[{"name":"IInd Department of Obstetrics Gynecology, Iuliu Ha\u0163ieganu University of Medicine and Pharmacy Cluj-Napoca, 13 Emil Isac, 400023 Cluj-Napoca, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mihaela","family":"Grigore","sequence":"additional","affiliation":[{"name":"Grigore T. Popa University of Medicine and Pharmacy, 700115 Iasi, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4718-392X","authenticated-orcid":false,"given":"Ingrid-Andrada","family":"Vasilache","sequence":"additional","affiliation":[{"name":"Grigore T. Popa University of Medicine and Pharmacy, 700115 Iasi, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1811-7923","authenticated-orcid":false,"given":"Ramona-Gabriela","family":"Ursu","sequence":"additional","affiliation":[{"name":"Grigore T. Popa University of Medicine and Pharmacy, 700115 Iasi, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Razvan","family":"Popovici","sequence":"additional","affiliation":[{"name":"Grigore T. Popa University of Medicine and Pharmacy, 700115 Iasi, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana-Maria","family":"Grigore","sequence":"additional","affiliation":[{"name":"Grigore T. Popa University of Medicine and Pharmacy, 700115 Iasi, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3994-6579","authenticated-orcid":false,"given":"Ludmila","family":"Lozneanu","sequence":"additional","affiliation":[{"name":"Grigore T. Popa University of Medicine and Pharmacy, 700115 Iasi, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-8427-8581","authenticated-orcid":false,"given":"Dan-Constantin","family":"Andronic","sequence":"additional","affiliation":[{"name":"Grigore T. Popa University of Medicine and Pharmacy, 700115 Iasi, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5374-0908","authenticated-orcid":false,"given":"Mitica","family":"Ciorpac","sequence":"additional","affiliation":[{"name":"Grigore T. Popa University of Medicine and Pharmacy, 700115 Iasi, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9240-2175","authenticated-orcid":false,"given":"Manuela","family":"Ciocoiu","sequence":"additional","affiliation":[{"name":"Grigore T. Popa University of Medicine and Pharmacy, 700115 Iasi, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,3,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Li, Y., Song, W., Gao, P., Guan, X., Wang, B., Zhang, L., Yao, Y., Guo, Y., Wang, Y., and Jiang, S. (2025). Global, regional, and national burden of breast, cervical, uterine, and ovarian cancer and their risk factors among women from 1990 to 2021, and projections to 2050: Findings from the global burden of disease study 2021. BMC Cancer, 25.","DOI":"10.1186\/s12885-025-13741-9"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"288","DOI":"10.1002\/ijc.35369","article-title":"Global landscape of cervical cancer incidence and mortality in 2022 and predictions to 2030: The urgent need to address inequalities in cervical cancer","volume":"157","author":"Li","year":"2025","journal-title":"Int. J. Cancer"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Bowden, S., Doulgeraki, T., Bouras, E., Markozannes, G., Athanasiou, A., Grout-Smith, H., Kechagias, K., Ellis, L., Zuber, V., and Chadeau-Hyam, M. (2023). Risk factors for human papillomavirus infection, cervical intraepithelial neoplasia and cervical cancer: An umbrella review and follow-up Mendelian randomisation studies. BMC Med., 21.","DOI":"10.1186\/s12916-023-02965-w"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"965","DOI":"10.1007\/s10147-023-02337-7","article-title":"Carcinogenesis and management of human papillomavirus-associated cervical cancer","volume":"28","author":"Kusakabe","year":"2023","journal-title":"Int. J. Clin. Oncol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Bautista, J., Altamirano-Colina, A., and L\u00f3pez-Cort\u00e9s, A. (2025). The vaginal microbiome in HPV persistence and cervical cancer progression. Front. Cell. Infect. Microbiol., 15.","DOI":"10.3389\/fcimb.2025.1634251"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1038\/s41564-022-01083-2","article-title":"Towards a deeper understanding of the vaginal microbiota","volume":"7","author":"France","year":"2022","journal-title":"Nat. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1186\/s12934-020-01464-4","article-title":"Vaginal microbiota and the potential of Lactobacillus derivatives in maintaining vaginal health","volume":"19","author":"Chee","year":"2020","journal-title":"Microb. Cell Fact."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1186\/s40168-022-01337-5","article-title":"Lactic acid from vaginal microbiota enhances cervicovaginal epithelial barrier integrity by promoting tight junction protein expression","volume":"10","author":"Jesaveluk","year":"2022","journal-title":"Microbiome"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1186\/s12934-025-02662-8","article-title":"Lactobacillus gasseri LGV03-derived indole-3-lactic acid ameliorates immune response by activating aryl hydrocarbon receptor","volume":"24","author":"Zhang","year":"2025","journal-title":"Microb. Cell Fact."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1186\/s40168-022-01400-1","article-title":"Vaginal microbiome-host interactions modeled in a human vagina-on-a-chip","volume":"10","author":"Mahajan","year":"2022","journal-title":"Microbiome"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Alizhan, D., Ukybassova, T., Bapayeva, G., Aimagambetova, G., Kongrtay, K., Kamzayeva, N., and Terzic, M. (2025). Cervicovaginal Microbiome: Physiology, Age-Related Changes, and Protective Role Against Human Papillomavirus Infection. J. Clin. Med., 14.","DOI":"10.3390\/jcm14051521"},{"key":"ref_12","first-page":"e85880","article-title":"Role of Vaginal and Gut Microbiota in Human Papillomavirus (HPV) Progression and Cervical Cancer: A Systematic Review of Microbial Diversity and Probiotic Interventions","volume":"17","author":"Pai","year":"2025","journal-title":"Cureus"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2449","DOI":"10.2147\/IJGM.S520936","article-title":"Analysis of the Relationship Between Cervical Cancer Progression and the Microbiome","volume":"18","author":"Gao","year":"2025","journal-title":"Int. J. Gen. Med."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Teka, B., Yoshida-Court, K., Firdawoke, E., Chanyalew, Z., Gizaw, M., Addissie, A., Mihret, A., Colbert, L., Napravnik, T., and Alam, M.E. (2023). Cervicovaginal Microbiota Profiles in Precancerous Lesions and Cervical Cancer among Ethiopian Women. Microorganisms, 11.","DOI":"10.3390\/microorganisms11040833"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Zhai, Q., Zhao, L., Wang, M., Li, L., Li, L., Ye, M., Li, M., Xu, C., and Meng, Y. (2025). Integrated analysis of microbiome and metabolome reveals insights into cervical neoplasia aggravation in a Chinese cohort. Front. Cell. Infect. Microbiol., 15.","DOI":"10.3389\/fcimb.2025.1556153"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1151","DOI":"10.1136\/ijgc-2020-001547","article-title":"Tumor microbial diversity and compositional differences among women in Botswana with high-grade cervical dysplasia and cervical cancer","volume":"30","author":"Sims","year":"2020","journal-title":"Int. J. Gynecol. Cancer"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1038\/s41392-025-02152-8","article-title":"Population-level analyses identify host and environmental variables influencing the vaginal microbiome","volume":"10","author":"Qin","year":"2025","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"11532","DOI":"10.1038\/s41598-024-62531-z","article-title":"Leveraging existing 16S rRNA gene surveys to decipher microbial signatures and dysbiosis in cervical carcinogenesis","volume":"14","author":"Li","year":"2024","journal-title":"Sci. Rep."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"816","DOI":"10.1186\/s12967-023-04676-5","article-title":"Characteristics of vaginal microbiota in various cervical intraepithelial neoplasia: A cross-sectional study","volume":"21","author":"Liu","year":"2023","journal-title":"J. Transl. Med."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"So, K.A., Yang, E.J., Kim, N.R., Hong, S.R., Lee, J.H., Hwang, C.S., Shim, S.H., Lee, S.J., and Kim, T.J. (2020). Changes of vaginal microbiota during cervical carcinogenesis in women with human papillomavirus infection. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0238705"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"348","DOI":"10.1016\/j.ygyno.2021.08.016","article-title":"The feature of cervical microbiota associated with the progression of cervical cancer among reproductive females","volume":"163","author":"Wu","year":"2021","journal-title":"Gynecol. Oncol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Wen, Q., Wang, S., Min, Y., Liu, X., Fang, J., Lang, J., and Chen, M. (2025). Associations of the gut, cervical, and vaginal microbiota with cervical cancer: A systematic review and meta-analysis. BMC Women\u2019s Health, 25.","DOI":"10.1186\/s12905-025-03599-1"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"e0019225","DOI":"10.1128\/spectrum.00192-25","article-title":"Microbial and metabolic profiles associated with HPV infection and cervical intraepithelial neoplasia: A multi-omics study","volume":"13","author":"Pu","year":"2025","journal-title":"Microbiol. Spectr."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1186\/s40246-025-00795-w","article-title":"Insights into the tripartite relationship between cervical cancer, human papillomavirus, and the vaginal microbiome: A mega-analysis","volume":"19","author":"Rashwan","year":"2025","journal-title":"Hum. Genom."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"e0267725","DOI":"10.1128\/mbio.02677-25","article-title":"The vaginal microbiome, papillomavirus infection, and cervical cancer: Established associations in search of model systems and mechanistic answers","volume":"2026","author":"Passarelli","year":"2026","journal-title":"mBio"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Schellekens, H.C.J., Schmidt, L.M.S., Morr\u00e9, S.A., van Esch, E.M.G., and de Vos van Steenwijk, P.J. (2025). Vaginal Microbiota and Local Immunity in HPV-Induced High-Grade Cervical Dysplasia: A Narrative Review. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26093954"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Lagunas-Cruz, M.D.C., Valle-Mendiola, A., and Soto-Cruz, I. (2026). The Vaginal Microbiome and Host Health: Implications for Cervical Cancer Progression. Int. J. Mol. Sci., 27.","DOI":"10.3390\/ijms27020640"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"767","DOI":"10.1093\/infdis\/jiae533","article-title":"Immunometabolic Contributions of Atopobiaceae Family Members in Human Papillomavirus Infection, Cervical Dysplasia, and Cancer","volume":"232","author":"Jimenez","year":"2025","journal-title":"J. Infect. Dis."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Chen, Y.-L., Qiu, X., Wang, W., Li, D., Wu, A., Hong, Z., Di, W., and Qiu, L. (2020). Human papillomavirus infection and cervical intraepithelial neoplasia progression are associated with increased vaginal microbiome diversity in a Chinese cohort. BMC Infect. Dis., 20.","DOI":"10.1186\/s12879-020-05324-9"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Mulato-Briones, I.B., Rodriguez-Ildefonso, I.O., Jim\u00e9nez-Tenorio, J.A., Cauich-S\u00e1nchez, P.I., M\u00e9ndez-Tovar, M.D.S., Aparicio-Ozores, G., Bautista-Hern\u00e1ndez, M.Y., Gonz\u00e1lez-Parra, J.F., Cruz-Hern\u00e1ndez, J., and L\u00f3pez-Romero, R. (2024). Cultivable Microbiome Approach Applied to Cervical Cancer Exploration. Cancers, 16.","DOI":"10.3390\/cancers16020314"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1999","DOI":"10.1038\/s41467-020-15856-y","article-title":"The vaginal microbiota associates with the regression of untreated cervical intraepithelial neoplasia 2 lesions","volume":"11","author":"Mitra","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"168","DOI":"10.1016\/j.trsl.2016.07.004","article-title":"Does the vaginal microbiota play a role in the development of cervical cancer?","volume":"179","author":"Kyrgiou","year":"2017","journal-title":"Transl. Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"987","DOI":"10.2147\/IJWH.S406596","article-title":"Imbalance of Vaginal Microbiota and Immunity: Two Main Accomplices of Cervical Cancer in Chinese Women","volume":"15","author":"Li","year":"2023","journal-title":"Int. J. Women\u2019s Health"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Kawasaki, R., Kukimoto, I., Nishio, E., Otani, S., Nishizawa, H., Maeda, Y., Iwata, A., and Fujii, T. (2025). Distinct cervical microbiome and metabolite profiles before and after menopause: Implications for cervical cancer progression. Front. Cell. Infect. Microbiol., 15.","DOI":"10.3389\/fcimb.2025.1589277"}],"container-title":["Diagnostics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4418\/16\/5\/753\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,3]],"date-time":"2026-03-03T15:48:53Z","timestamp":1772552933000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4418\/16\/5\/753"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,3]]},"references-count":34,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2026,3]]}},"alternative-id":["diagnostics16050753"],"URL":"https:\/\/doi.org\/10.3390\/diagnostics16050753","relation":{},"ISSN":["2075-4418"],"issn-type":[{"value":"2075-4418","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,3]]}}}