Human papilloma virus testing in oropharyngeal squamous cell carcinoma: what the clinician should know

Oral Oncol. 2014 Jan;50(1):1-9. doi: 10.1016/j.oraloncology.2013.10.008. Epub 2013 Oct 26.


High risk Human Papilloma virus (HR-HPV) associated oropharyngeal cancers are on the increase. Although, the scientific community is aware of the importance of Human Papilloma Virus (HPV) testing, there is no consensus on the assays that are required to reliably identify HR-HPV related tumors. A wide range of methods have been developed. The most widely used techniques include viral DNA detection, with polymerase chain reaction (PCR) or In Situ Hybridization, and p16 detected by immunohistochemistry. However, these tests provide different information and have their own specific limitations. In this review, we summarize these different techniques, in light of the recent literature. p16 Overexpression, which is an indirect marker of HPV infection, is considered by many head and neck oncologists to be the most important marker for patient stratification. We describe the frequent lack of concordance of this marker with other assays and the possible reasons for this. The latest developments in HPV testing are also reported, such as the RNAscope™ HPV test, and how they fit into the existing framework of techniques. HPV testing must not be considered in isolation, as there are important interactions with other parameters, such as tobacco exposure. This is an important and rapidly evolving field and is likely to become pivotal to staging and choice of treatment of oropharyngeal carcinoma in the future.

Keywords: HPV; Head and neck cancer; Molecular diagnosis; Oropharyngeal cancer; p16.

Publication types

  • Review

MeSH terms

  • Carcinoma, Squamous Cell / virology*
  • DNA, Viral / analysis
  • Humans
  • In Situ Hybridization
  • Oropharyngeal Neoplasms / virology*
  • Papillomaviridae / genetics
  • Papillomaviridae / isolation & purification
  • Papillomavirus Infections / diagnosis*
  • Polymerase Chain Reaction
  • RNA, Messenger / analysis


  • DNA, Viral
  • RNA, Messenger