title: Oral swirl samples - a robust source of microRNA protected by extracellular vesicles.
abstract: BACKGROUND
  MicroRNAs are small non-coding RNAs which are dysregulated in disease states, such as oral cancer. Extracellular vesicles, a potential source of microRNA, are found in saliva.
  
  
  OBJECTIVE
  To demonstrate that a quantifiable amount of microRNA can be isolated from oral swirl samples. Additionally, we hypothesized that extracellular vesicles may protect contained microRNA from degradation in these samples.
  
  
  METHODS
  A polyethylene glycol-based precipitation was used for extracellular vesicle enrichment of oral swirl samples. Comparison was made between samples treated with and without RNase. Further, samples from three subjects were exposed to a range of conditions over 7 days and assessed for presence of microRNA by reverse-transcription quantitative PCR. Extracellular vesicles from samples were identified under transmission electron microscopy.
  
  
  RESULTS
  An adequate quantity of microRNA for qPCR analysis was extractable from samples despite exposure to conditions under which degradation of RNA would be expected.
  
  
  CONCLUSIONS
  A technique was developed to isolate an adequate quantity of microRNA for analysis from oral swirl samples. Extracellular vesicle-associated microRNA may be protected from degradation. This technique moves towards chairside application of translational microRNA research in the field of oral cancer prognostics.
authors: Melbourne Dental School, University of Melbourne, Melbourne, Vic., Australia.
authors: T
authors: T
authors: Yap
authors: The Florey Institute of Neuroscience and Mental Health, Parkville, Vic., Australia.
authors: L J
authors: LJ
authors: Vella
authors: Melbourne Dental School, University of Melbourne, Melbourne, Vic., Australia.|Oral Health Cooperative Research Centre, Melbourne, Vic., Australia.
authors: C
authors: C
authors: Seers
authors: Department of Oral and Maxillofacial Surgery, Royal Melbourne Hospital, Melbourne, Vic., Australia.
authors: A
authors: A
authors: Nastri
authors: Melbourne Dental School, University of Melbourne, Melbourne, Vic., Australia.|Oral Health Cooperative Research Centre, Melbourne, Vic., Australia.
authors: E
authors: E
authors: Reynolds
authors: Melbourne Dental School, University of Melbourne, Melbourne, Vic., Australia.|Oral Health Cooperative Research Centre, Melbourne, Vic., Australia.
authors: N
authors: N
authors: Cirillo
authors: Melbourne Dental School, University of Melbourne, Melbourne, Vic., Australia.|Oral Health Cooperative Research Centre, Melbourne, Vic., Australia.
authors: M
authors: M
authors: McCullough
mesh_terms: D000067128:Extracellular Vesicles; D006801:Humans; D035683:MicroRNAs / Q000302:isolation & purification*; D009055:Mouth; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012463:Saliva / Q000737:chemistry*; D013048:Specimen Handling / Q000379:methods*
chemical_list: D035683:MicroRNAs
keywords: extracellular vesicles; microRNA; oral cancer; saliva