Abstract
Evidence suggests the vaginal microbiota (VM) may influence risk of persistent Human Papillomavirus (HPV) infection and cervical carcinogenesis. Established cytology biobanks, typically collected with a cytobrush, constitute a unique resource to study such associations longitudinally. It is plausible that compared to rayon swabs; the most commonly used sampling devices, cytobrushes may disrupt biofilms leading to variation in VM composition. Cervico-vaginal samples were collected with cytobrush and rayon swabs from 30 women with high-grade cervical precancer. Quantitative PCR was used to compare bacterial load and Illumina MiSeq sequencing of the V1-V3 regions of the 16S rRNA gene used to compare VM composition. Cytobrushes collected a higher total bacterial load. Relative abundance of bacterial species was highly comparable between sampling devices (R2 = 0.993). However, in women with a Lactobacillus-depleted, high-diversity VM, significantly less correlation in relative species abundance was observed between devices when compared to those with a Lactobacillus species-dominant VM (p = 0.0049). Cytobrush and swab sampling provide a comparable VM composition. In a small proportion of cases the cytobrush was able to detect underlying high-diversity community structure, not realized with swab sampling. This study highlights the need to consider sampling devices as potential confounders when comparing multiple studies and datasets.
Highlights
Cervical cancer is a disease that has become largely preventable thanks to screening programmes that allow detection and treatment of pre-invasive disease[1]
As estimated using quantitative PCR, cytobrushes collected a greater total bacterial load when compared to swabs (Fig. 1a, Table 1)
Cross-sectional studies exploring the associations between the vaginal microbiota (VM), Human Papillomavirus (HPV) infection and cervical pre-invasive and invasive disease have shown that a high-diversity VM, and to a lesser extent L. iners-dominant VM’s correlate with increasing cervical disease severity[7, 8, 10], and in acquisition and persistence of its causative agent HPV5
Summary
Cervical cancer is a disease that has become largely preventable thanks to screening programmes that allow detection and treatment of pre-invasive disease (cervical intraepithelial neoplasia; CIN)[1]. A two-group comparison of the different sampling techniques was performed showing that relative abundance of bacterial taxa was highly comparable between all swabs and cytobrushes (R2 = 0.998–0.999 from phylum to genus level, R2 = 0.993 at species level) (Fig. 5A).
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