Shende Smriti, Das Rashmita, Joshi Suvarna, Yanamandra Sushma, Taji Nyabom, Karyakarte Rajesh P
Microbiology, B. J. Government Medical College & Sassoon General Hospitals, Pune, IND.
Microbiology, Tomo Riba Institute of Health and Medical Sciences, Itanagar, IND.
Cureus. 2025 Aug 12;17(8):e89872. doi: 10.7759/cureus.89872. eCollection 2025 Aug.
Cervical cancer is a common cancer among women worldwide, especially in low- and middle-income countries. Persistent infection with high-risk human papillomavirus (HR-HPV) largely drives the development of cervical cancer. While -dominant communities are considered protective, dysbiosis, marked by reduced and increased anaerobic diversity (community state type (CST)-IV), may promote viral persistence. This study aims to assess HR-HPV prevalence and compare vaginal microbiome profiles in women with suspected HPV infection and healthy controls.
Vaginal swabs were collected from pre-menopausal women with clinically suspected HPV infection and healthy controls. Samples underwent HR-HPV detection using the Truenat® HPV-HR RT-PCR (real-time polymerase chain reaction) assay (Molbio Diagnostics Limited, Goa, India). Vaginal microbiome profiling was performed using 16S rRNA gene amplicon sequencing on the GridION platform (Oxford Nanopore Technologies plc, Oxford, United Kingdom). Taxonomic classification was carried out using the EPI2ME 16S workflow with Kraken2 (Johns Hopkins University, Baltimore, Maryland, United States), and CSTs were assigned using the VALENCIA (VAginaL community state typE Nearest CentroId clAssifier) algorithm. Statistical analyses and microbial community comparisons were performed using MicrobiomeAnalyst (Xia Lab, Sainte-Anne-de-Bellevue, Quebec, Canada).
A total of 86 clinically suspected HPV cases and 63 healthy pre-menopausal controls were enrolled. Overall, high-risk HPV (HPV 16/31 and HPV-18/45) was detected in 11.6% (10/86) of cases, with no positivity in the control group. Vaginal microbiome profiling revealed significantly higher alpha diversity in women with inflammatory cytology compared to healthy controls, and increased Shannon and Simpson diversity indices in HPV-positive and inflammatory groups. Beta diversity analysis showed distinct microbial clustering between all groups. Taxonomic analysis demonstrated a predominance of spp. in healthy individuals, particularly and , whereas disease groups showed increased abundance of , , and . CST-I was more common in healthy controls (7/17, 41.2%), while CST-IV-linked to dysbiosis-was predominant among suspected cases (9/21, 47.6%). Linear discriminant analysis effect size (LEfSe) analysis identified , , and as key genera in healthy individuals, whereas , , and were enriched in HPV-positive and inflammatory smear groups, highlighting potential microbial biomarkers of vaginal dysbiosis.
These findings underscore the importance of vaginal microbiome composition in cervical health and support further investigation into microbial biomarkers for early detection and targeted interventions in HPV-associated disease.
宫颈癌是全球女性中的常见癌症，尤其是在低收入和中等收入国家。高危型人乳头瘤病毒（HR-HPV）的持续感染在很大程度上推动了宫颈癌的发展。虽然以乳酸杆菌为主的群落被认为具有保护作用，但以乳酸杆菌减少和厌氧多样性增加为特征的生态失调（群落状态类型（CST）-IV）可能会促进病毒持续存在。本研究旨在评估疑似HPV感染女性和健康对照者中HR-HPV的患病率，并比较其阴道微生物群谱。
从临床疑似HPV感染的绝经前女性和健康对照者中采集阴道拭子。使用Truenat®HPV-HR RT-PCR（实时聚合酶链反应）检测法（印度果阿Molbio诊断有限公司）对样本进行HR-HPV检测。在GridION平台（英国牛津牛津纳米孔技术有限公司）上使用16S rRNA基因扩增子测序进行阴道微生物群分析。使用带有Kraken2的EPI2ME 16S工作流程（美国马里兰州巴尔的摩约翰霍普金斯大学）进行分类学分类，并使用VALENCIA（阴道群落状态类型最近质心分类器）算法指定CST。使用微生物组分析软件（加拿大魁北克省圣安妮-德-贝尔维尤夏实验室）进行统计分析和微生物群落比较。
共纳入86例临床疑似HPV病例和63例健康绝经前对照者。总体而言，11.6%（10/86）的病例检测到高危型HPV（HPV 16/31和HPV-18/45），对照组均为阴性。阴道微生物群分析显示，与健康对照者相比，炎症细胞学女性的α多样性显著更高，HPV阳性组和炎症组的香农和辛普森多样性指数增加。β多样性分析显示所有组之间存在明显的微生物聚类。分类学分析表明，健康个体中以卷曲乳酸杆菌属为主，尤其是卷曲乳酸杆菌和詹氏乳酸杆菌，而疾病组中惰性乳杆菌、加氏乳杆菌和卷曲乳杆菌的丰度增加。CST-I在健康对照者中更为常见（7/17，41.2%），而与生态失调相关的CST-IV在疑似病例中占主导地位（9/21，47.6%）。线性判别分析效应大小（LEfSe）分析确定卷曲乳酸杆菌、詹氏乳酸杆菌和嗜酸乳杆菌为健康个体中的关键属，而惰性乳杆菌、加氏乳杆菌和卷曲乳杆菌在HPV阳性和炎症涂片组中富集，突出了阴道生态失调的潜在微生物生物标志物。
这些发现强调了阴道微生物群组成在宫颈健康中的重要性，并支持进一步研究微生物生物标志物，以早期检测和针对性干预HPV相关疾病。
