Volpi CC, et al. - As a part of this study, two in situ hybridization (ISH) detection methods for human papilloma virus (HPV) 16 E6/E7 mRNA, i.e. the RNAscope 2.0 High Definition (HD) and the upgraded RNAscope 2.5 HD version, underwent a comparative analysis. The data brought to the limelight was that the compared RNAscope HPV tests displayed a good level of interchangeability and facilitated the detection of oropharyngeal squamous cell carcinoma (OSCC), driven by high risk-HPV infection. It gained support via the comparison of the RNAscope HPV test with HPV E6/E7 mRNA real-time reverse transcriptase-polymerase chain reaction (RT-PCR), in a fraction of cases where the material for HPV E6/E7 mRNA real-time RT-PCR was available. The algorithm correlating with p16 immunohistochemistry (IHC) alongside the detection of HPV mRNA by RNAscope was unveiled to be more effective than the one that related p16 IHC with the identification of HPV DNA by ISH.