Author Identifier (ORCID)
Claus T. Christophersen: https://orcid.org/0000-0003-1591-5871
Abstract
Vaginal microbiome composition has been linked to risk of preterm birth (PTB), a persistent global health challenge. 16S rRNA microbial profiling has identified specific vaginal community state types (CSTs) that have been associated with PTB risk. Diagnostic profiling requires standardised pre-analytical protocols. We evaluated two storage methods and validated a curated, vagina-specific 16S rRNA gene database (VagDB) to enhance annotation. Paired Copan FLOQ swabs from 22 women at high PTB risk were processed for either (a) dry/immediate freezing or (b) Amies-stabilisation/refrigeration. Amplicon sequence variants were generated via 16S rRNA gene (V4) PCR and Illumina sequencing. We assessed diversity, composition, and community state type (CST) allocation. Amies-stabilised samples yielded significantly higher DNA (p = 0.003), but this did not alter species richness, evenness, or community structure. VagDB enhanced species-level resolution. PCoA showed robust clustering by participant and CST (p < 0.001), irrespective of storage; CST concordance exceeded 90%. Routinely collected vaginal swabs in stabilisation medium with an 8–72 h refrigeration window yield reliable data, supporting the integration of vaginal microbiome profiling into clinical PTB risk assessment.
Keywords
16S rRNA, amplicon sequencing, community state types, preterm birth, swab sampling, vaginal database, vaginal microbiome
Document Type
Journal Article
Date of Publication
1-1-2026
Volume
14
Issue
1
Publication Title
Microorganisms
Publisher
MDPI
School
School of Medical and Health Sciences
Funders
Western Australian Human Microbiome Collaboration Centre (WAHMCC) / Curtin University / Channel 7 Telethon Trust / Western Australia Pregnancy Biobank / Women & Infants Research Foundation
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.
Comments
Ali, A., Keelan, J. A., Penova-Veselinovic, B., Allentoft, M. E., Bunce, M., & Christophersen, C. T. (2026). Optimising vaginal microbiome profiling for clinical translation: A comparative assessment of sample storage methods and a vagina-specific 16s rRNA gene database. Microorganisms, 14(1). https://doi.org/10.3390/microorganisms14010128