Comparative analysis of vaginal microbiota sampling using 16S rRNA gene analysis

Area of Science:

• Microbiome research
• Gynecological health
• Molecular diagnostics

Background:

• Vaginal microbiota characterization is crucial for understanding health and disease.
• Previous research primarily focused on biological variations, with less attention to sampling method impacts.
• Understanding how sampling impacts results is vital for reliable vaginal microbiota studies.

Purpose of the Study:

• To investigate the influence of sampling devices and anatomical sites on vaginal microbiota analysis.
• To compare quantitative (DNA/protein yield, bacterial load) and qualitative (diversity, composition) outcomes.
• To determine the relative contributions of sampling versus intrinsic factors to microbiota variation.

Main Methods:

• 10 women provided samples using flocked swabs, Evalyn® self-sampling device, spatulas, and a cervical brush.
• Samples were collected from the fornix, vaginal wall, and cervix.
• 16S rRNA gene sequencing on Illumina MiSeq platform analyzed microbiota diversity and composition.

Main Results:

• Sampling device significantly affected DNA yield, but not protein yield or bacterial load.
• Microbiota profiles were comparable across all tested sampling methods.
• Intrinsic factors (clinical status, bleeding, individual variation) explained most microbiota variation (83%), while sampling method explained only 2%.

Conclusions:

• Different vaginal microbiota sampling strategies yield comparable results, ensuring consistency across studies.
• The Evalyn® self-sampling device provides high-quality samples comparable to clinician-collected ones, enabling non-invasive collection.
• While sampling techniques vary in sample/yield amounts, they do not compromise overall microbiota comparability.