title: The effect of propidium monoazide treatment on the measured bacterial composition of clinical samples after the use of a mouthwash.
abstract: OBJECTIVE
  The use of an anti-microbial mouthwash results not only in a reduction of the number of viable cells in dental plaque but potentially also in a shift in the oral microbiome. DNA-based techniques may be appropriate to monitor these shifts, but these techniques amplify DNA from both dead and living cells. Propidium monoazide (PMA) has been used to overcome this problem, by preventing the amplification of DNA from membrane-damaged cells. The aim of this study was to evaluate the use of PMA when measuring compositional shifts in clinical samples after mouthwash use.
  
  
  METHODS
  On two consecutive days, baseline samples from buccal surfaces, tongue, and saliva were obtained from six volunteers, after which they used a mouthwash (Meridol, GABA, Switzerland) twice daily for 14 days. Subsequently similar samples were obtained on two consecutive days. The microbial composition of the samples, with or without ex vivo PMA treatment, was assessed with 16S rRNA gene amplicon sequencing.
  
  
  RESULTS
  Data showed a clear effect of mouthwash usage on the tongue and saliva samples. PMA treatment enhanced the observed differences only for the saliva samples. Mouthwash treatments did not affect the composition of the plaque samples irrespective of the use of PMA.
  
  
  CONCLUSIONS
  The necessity to use a PMA treatment to block the DNA from dead cells in clinical studies aimed at measuring compositional shifts after the use of a mouthwash is limited to salivary samples.
  
  
  CONCLUSIONS
  Measuring shifts in the oral microbiome could be hampered by the presence of DNA from dead cells.
authors.affiliation: Department of Preventive Dentistry, Academic Centre for Dentistry Amsterdam (ACTA), University of Amsterdam and VU University Amsterdam, Amsterdam, The Netherlands, R.Exterkate@acta.nl.
authors.forename: R A M
authors.initials: RA
authors.lastname: Exterkate
authors.forename: E
authors.initials: E
authors.lastname: Zaura
authors.forename: B W
authors.initials: BW
authors.lastname: Brandt
authors.forename: M J
authors.initials: MJ
authors.lastname: Buijs
authors.forename: J E
authors.initials: JE
authors.lastname: Koopman
authors.forename: W
authors.initials: W
authors.lastname: Crielaard
authors.forename: J M
authors.initials: JM
authors.lastname: Ten Cate
mesh_terms: D001386:Azides / Q000737:chemistry / Q000494:pharmacology*; D004269:DNA, Bacterial; D003773:Dental Plaque / Q000382:microbiology; D006801:Humans; D064307:Microbiota / Q000187:drug effects*; D009067:Mouthwashes / Q000737:chemistry / Q000494:pharmacology*; D025341:Principal Component Analysis; D011419:Propidium / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D012463:Saliva / Q000382:microbiology*
chemical_list: D001386:Azides; D004269:DNA, Bacterial; D009067:Mouthwashes; C533957:propidium monoazide; D011419:Propidium
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