Validation of a novel diagnostic standard in HPV-positive oropharyngeal squamous cell carcinoma

A G Schache, T Liloglou, J M Risk, T M Jones, X-J Ma, H Wang, S Bui, Y Luo, P Sloan, R J Shaw, M Robinson

Research output: Contribution to journalArticle (journal)peer-review

144 Citations (Scopus)


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.

CONCLUSION: 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.

Original languageEnglish
Pages (from-to)1332-9
Number of pages8
JournalBritish Journal of Cancer
Issue number6
Publication statusPublished - 2 Apr 2013


  • Biomarkers, Tumor/genetics
  • Carcinoma, Squamous Cell/diagnosis
  • Female
  • Follow-Up Studies
  • Humans
  • Male
  • Middle Aged
  • Molecular Diagnostic Techniques
  • Neoplasm Staging
  • Oropharyngeal Neoplasms/diagnosis
  • Papillomaviridae/genetics
  • Papillomavirus Infections/diagnosis
  • Polymerase Chain Reaction
  • Prognosis
  • RNA, Viral/genetics
  • Survival Rate
  • Tissue Array Analysis


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