Characterization of vaginal microbiota diversity by 16S rRNA high-throughput sequencing.
Front Microbiol 2026; 17:1777216.

Abstract

Introduction

This study employed high-throughput sequencing to analyze the diversity of vaginal microbiota in healthy women of reproductive age, thereby establishing a foundational dataset that may inform future forensic applications such as individual identification.

Methods

A cohort of 100 healthy reproductive-age women meeting the inclusion criteria was enrolled. Vaginal secretion samples were collected, after which microbial DNA was extracted and assessed for quality using agarose gel electrophoresis. The V3-V4 region of the 16S rRNA gene was subsequently amplified with universal primers and subjected to Illumina high-throughput sequencing for comprehensive microbial analysis.

Results

Sequencing analysis revealed that the five most predominant genera in the female vaginal microbiota were Lactobacillus, Gardnerella, Prevotella, Serratia, and Atopobium. Cluster analysis of the 100 samples classified them into five major clusters, further delineated into eight subtypes, ranging from absolute dominance of Lactobacillus to dominance of non-Lactobacillus bacteria. Analysis of influence factors showed that microbial diversity was positively associated with age, higher in urban compared to suburban participants, and lower in pregnant versus non-pregnant women. The 16S rRNA high-throughput sequencing of vaginal microbiota exhibited distinct inter-group clustering, with Lactobacillus identified as a core candidate marker for vaginal secretions.

Discussion

The compositional profile of the vaginal microbiota exhibited variations associated with individual characteristics such as age range and geographic region. This study provides a foundational description of vaginal microbiota composition in this cohort. Such foundational data are a necessary first step toward exploring potential future applications in forensic science, though direct inference of individual characteristics is not supported by the current dataset.

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Authors+Show Affiliations

Chen MLuoyang Key Laboratory of Zebrafish Toxicology Research, Luoyang Key Laboratory of Transplantation and Immunological Studies for Haematological Diseases, Department of Clinical Laboratory, the First Affiliated Hospital, College of Clinical Medicine of Henan University of Science and Technology, Luoyang, China.
Li JSchool of Basic Medicine and Forensic Medicine, Henan University of Science and Technology, Luoyang, China. Criminal Investigation Brigade of Xuanwu Sub-bureau, Nanjing Public Security Bureau, Nanjing, China.
Chen HLuoyang Key Laboratory of Zebrafish Toxicology Research, Luoyang Key Laboratory of Transplantation and Immunological Studies for Haematological Diseases, Department of Clinical Laboratory, the First Affiliated Hospital, College of Clinical Medicine of Henan University of Science and Technology, Luoyang, China.
Tuerxun ASchool of Basic Medicine and Forensic Medicine, Henan University of Science and Technology, Luoyang, China.
Mao MSchool of Basic Medicine and Forensic Medicine, Henan University of Science and Technology, Luoyang, China.
Song JSchool of Basic Medicine and Forensic Medicine, Henan University of Science and Technology, Luoyang, China.
Li BSchool of Basic Medicine and Forensic Medicine, Henan University of Science and Technology, Luoyang, China.
Mo YSchool of Basic Medicine and Forensic Medicine, Henan University of Science and Technology, Luoyang, China.
Zhai XSchool of Basic Medicine and Forensic Medicine, Henan University of Science and Technology, Luoyang, China.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

42088275