title: Validation of a novel diagnostic standard in HPV-positive oropharyngeal squamous cell carcinoma.
abstract: BACKGROUND
  Human papillomavirus (HPV) testing in oropharyngeal squamous cell carcinoma (OPSCC) is now advocated. Demonstration of transcriptionally active high-risk HPV (HR-HPV) in fresh tumour tissue is considered to be the analytical 'gold standard'. Clinical testing has focused on formalin-fixed paraffin-embedded (FFPE) tissue at the expense of sensitivity and specificity. Recently, a novel RNA in situ hybridisation test (RNAscope) has been developed for the detection of HR-HPV in FFPE tissue; however, validation against the 'gold standard' has not been reported.
  
  
  METHODS
  A tissue microarray comprising FFPE cores from 79 OPSCC was tested using HR-HPV RNAscope. Analytical accuracy and prognostic capacity were established by comparison with the reference test; qRT-PCR for HR-HPV on matched fresh-frozen samples.
  
  
  RESULTS
  High-risk HPV RNAscope had a sensitivity and specificity of 97 and 93%, respectively, against the reference test. Kaplan-Meier estimates of disease-specific survival (DSS, P=0.001) and overall survival (OS, P<0.001) by RNAscope were similar to the reference test (DSS, P=0.003, OS, P<0.001) and at least, not inferior to p16 immunohistochemistry +/- HR-HPV DNA-based tests.
  
  
  CONCLUSIONS
  HR-HPV RNAscope demonstrates excellent analytical and prognostic performance against the 'gold standard'. These data suggest that the test could be developed to provide the 'clinical standard' for assigning a diagnosis of HPV-related OPSCC.
authors: Department of Molecular and Clinical Cancer Medicine, Institute of Translational Medicine, University of Liverpool, 3.01 Research Wing, Daulby Street, Liverpool L69 3GN, UK. schache@liverpool.ac.uk
authors: A G
authors: AG
authors: Schache
authors: T
authors: T
authors: Liloglou
authors: J M
authors: JM
authors: Risk
authors: T M
authors: TM
authors: Jones
authors: X-J
authors: XJ
authors: Ma
authors: H
authors: H
authors: Wang
authors: S
authors: S
authors: Bui
authors: Y
authors: Y
authors: Luo
authors: P
authors: P
authors: Sloan
authors: R J
authors: RJ
authors: Shaw
authors: M
authors: M
authors: Robinson
mesh_terms: D014408:Biomarkers, Tumor / Q000235:genetics*; D002294:Carcinoma, Squamous Cell / Q000175:diagnosis* / Q000401:mortality / Q000821:virology; D005260:Female; D005500:Follow-Up Studies; D006801:Humans; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques; D009367:Neoplasm Staging; D009959:Oropharyngeal Neoplasms / Q000175:diagnosis* / Q000401:mortality / Q000821:virology; D027383:Papillomaviridae / Q000235:genetics*; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000401:mortality / Q000821:virology; D016133:Polymerase Chain Reaction; D011379:Prognosis; D012367:RNA, Viral / Q000235:genetics; D015996:Survival Rate; D046888:Tissue Array Analysis
chemical_list: D014408:Biomarkers, Tumor; D012367:RNA, Viral