Saliva sampling method influences oral microbiome composition and taxa distribution associated with oral diseases

PLoS One. 2024 Mar 28;19(3):e0301016. doi: 10.1371/journal.pone.0301016. eCollection 2024.

Abstract

Saliva is a readily accessible and inexpensive biological specimen that enables investigation of the oral microbiome, which can serve as a biomarker of oral and systemic health. There are two routine approaches to collect saliva, stimulated and unstimulated; however, there is no consensus on how sampling method influences oral microbiome metrics. In this study, we analyzed paired saliva samples (unstimulated and stimulated) from 88 individuals, aged 7-18 years. Using 16S rRNA gene sequencing, we investigated the differences in bacterial microbiome composition between sample types and determined how sampling method affects the distribution of taxa associated with untreated dental caries and gingivitis. Our analyses indicated significant differences in microbiome composition between the sample types. Both sampling methods were able to detect significant differences in microbiome composition between healthy subjects and subjects with untreated caries. However, only stimulated saliva revealed a significant association between microbiome diversity and composition in individuals with diagnosed gingivitis. Furthermore, taxa previously associated with dental caries and gingivitis were preferentially enriched in individuals with each respective disease only in stimulated saliva. Our study suggests that stimulated saliva provides a more nuanced readout of microbiome composition and taxa distribution associated with untreated dental caries and gingivitis compared to unstimulated saliva.

MeSH terms

  • Dental Caries*
  • Gingivitis*
  • Humans
  • Microbiota* / genetics
  • RNA, Ribosomal, 16S / genetics
  • Saliva / microbiology

Substances

  • RNA, Ribosomal, 16S

Grants and funding

C.R., D.L.C and M.C.W. were funded in part by support from the National Institute of Dental and Craniofacial Research, NIH (U01DE030418), and J.R.W. was funded by NIH K01 DK119582. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.