Methodological approaches in 16S sequencing of female reproductive tract in fertility patients: a review

Izzy Davidson, Elham Nikbakht Nasrabadi, Larisa M Haupt, Kevin John Ashton, Paul J Dunn*

*Corresponding author for this work

Research output: Contribution to journalReview articleResearchpeer-review

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Abstract

Background:
The female genital tract microbiome has become a particular area of interest in improving assisted reproductive technology (ART) outcomes with the emergence of next-generation sequencing (NGS) technology. However, NGS assessment of microbiomes currently lacks uniformity and poses signifcant challenges for accurate and precise bacterial population representation.

Objective:
As multiple NGS platforms and assays have been developed in recent years for microbiome investigation—including the advent of long-read sequencing technologies—this work aimed to identify current trends and practices undertaken in female genital tract microbiome investigations.

Results:
Areas like sample collection and transport, DNA extraction, 16S amplifcation vs. metagenomics, NGS library preparation, and bioinformatic analysis demonstrated a detrimental lack of uniformity. The lack of uniformity present is a signifcant limitation characterised by gap discrepancies in generation and interpretation of results. Minimal consistency was observed in primer design, DNA extraction techniques, sample transport, and bioinformatic analyses.

Conclusion:
With third-generation sequencing technology highlighted as a promising tool in microbiota-based research via full-length 16S rRNA sequencing, there is a desperate need for future studies to investigate and optimise methodological approaches of the genital tract microbiome to ensure better uniformity of methods and results interpretation to improve
clinical impact.
Original languageEnglish
Pages (from-to)1-23
Number of pages23
JournalJournal of Assisted Reproduction and Genetics
DOIs
Publication statusPublished - 21 Oct 2024

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