title: A pilot study to compare the detection of HPV-16 biomarkers in salivary oral rinses with tumour p16(INK4a) expression in head and neck squamous cell carcinoma patients.
abstract: BACKGROUND
  Human papilloma virus-16 (HPV-16) infection is a major risk factor for a subset of head and neck squamous cell carcinoma (HNSCC), in particular oropharyngeal squamous cell carcinoma (OPSCC). Current techniques for assessing the HPV-16 status in HNSCC include the detection of HPV-16 DNA and p16(INK4a) expression in tumor tissues. When tumors originate from hidden anatomical sites, this method can be challenging. A non-invasive and cost-effective alternative to biopsy is therefore desirable for HPV-16 detection especially within a community setting to screen at-risk individuals.
  
  
  METHODS
  The present study compared detection of HPV-16 DNA and RNA in salivary oral rinses with tumor p16(INK4a) status, in 82 HNSCC patients using end-point and quantitative polymerase chain reaction (PCR).
  
  
  RESULTS
  Of 42 patients with p16(INK4a)-positive tumours, 39 (sensitivity = 92.9 %, PPV = 100 % and NPV = 93 %) had oral rinse samples with detectable HPV-16 DNA, using end-point and quantitative PCR. No HPV-16 DNA was detected in oral rinse samples from 40 patients with p16(INK4a) negative tumours, yielding a test specificity of 100 %. For patients with p16(INK4a) positive tumours, HPV-16 mRNA was detected using end-point reverse transcription PCR (RT-PCR) in 24/40 (sensitivity = 60 %, PPV = 100 % and NPV = 71 %), and using quantitative RT-PCR in 22/40 (sensitivity = 55 %, PPV = 100 % and NPV = 69 %). No HPV-16 mRNA was detected in oral rinse samples from the p16(INK4a)-negative patients, yielding a specificity of 100 %.
  
  
  CONCLUSIONS
  We demonstrate that the detection of HPV-16 DNA in salivary oral rinse is indicative of HPV status in HNSCC patients and can potentially be used as a diagnostic tool in addition to the current methods.
authors.affiliation: The University of Queensland Diamantina Institute, The University of Queensland, Translational Research Institute, 37 Kent St, Woolloongabba, QLD, Australia. r.chai@garvan.org.au.|Present address: Garvan Institute of Medical Research, 384 Victoria St, Darlinghurst, NSW, Australia. r.chai@garvan.org.au.
authors.forename: Ryan C
authors.initials: RC
authors.lastname: Chai
authors.affiliation: The School of Biomedical Sciences, Institute of Health and Biomedical Innovations, Queensland University of Technology, 60 Musk Avenue, Kelvin Grove, QLD, Australia. y43.lim@qut.edu.au.
authors.forename: Yenkai
authors.initials: Y
authors.lastname: Lim
authors.affiliation: The University of Queensland Diamantina Institute, The University of Queensland, Translational Research Institute, 37 Kent St, Woolloongabba, QLD, Australia. i.frazer@uq.edu.au.
authors.forename: Ian H
authors.initials: IH
authors.lastname: Frazer
authors.affiliation: The School of Biomedical Sciences, Institute of Health and Biomedical Innovations, Queensland University of Technology, 60 Musk Avenue, Kelvin Grove, QLD, Australia. yunxia.wan@qut.edu.au.
authors.forename: Yunxia
authors.initials: Y
authors.lastname: Wan
authors.affiliation: Princess Alexandra Hospital, 199 Ipswich Rd, Woolloongabba, QLD, Australia. reception@brisbaneent.com.au.
authors.forename: Christopher
authors.initials: C
authors.lastname: Perry
authors.affiliation: The School of Biomedical Sciences, Institute of Health and Biomedical Innovations, Queensland University of Technology, 60 Musk Avenue, Kelvin Grove, QLD, Australia. lee.jones@qut.edu.au.
authors.forename: Lee
authors.initials: L
authors.lastname: Jones
authors.affiliation: IQ Pathology, 6/11 Donkin St, West End, QLD, Australia. d.lambie@uq.edu.au.
authors.forename: Duncan
authors.initials: D
authors.lastname: Lambie
authors.affiliation: The School of Biomedical Sciences, Institute of Health and Biomedical Innovations, Queensland University of Technology, 60 Musk Avenue, Kelvin Grove, QLD, Australia. chamindie.punyadeera@qut.edu.au.
authors.forename: Chamindie
authors.initials: C
authors.lastname: Punyadeera
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor; D002294:Carcinoma, Squamous Cell / Q000209:etiology*; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics*; D004279:DNA, Viral; D005260:Female; D005814:Genes, Viral; D006258:Head and Neck Neoplasms / Q000209:etiology*; D052162:Human papillomavirus 16 / Q000235:genetics*; D006801:Humans; D008297:Male; D008875:Middle Aged; D030361:Papillomavirus Infections / Q000150:complications* / Q000821:virology; D016133:Polymerase Chain Reaction; D012463:Saliva; D000077195:Squamous Cell Carcinoma of Head and Neck
chemical_list: D014408:Biomarkers, Tumor; D019941:Cyclin-Dependent Kinase Inhibitor p16; D004279:DNA, Viral