In situ hybridization detection methods for HPV16 E6/E7 mRNA in identifying transcriptionally active HPV infection of oropharyngeal carcinoma: an updating

Publication date: Available online 6 October 2017
Source:Human Pathology
Author(s): Chiara C. Volpi, Chiara M. Ciniselli, Ambra V. Gualeni, Maddalena Plebani, Salvatore Alfieri, Paolo Verderio, Laura Locati, Federica Perrone, Pasquale Quattrone, Antonino Carbone, Silvana Pilotti, Annunziata Gloghini
The aim of this study is comparing 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. The RNAscope® 2.5 HD has recently replaced the RNAscope® 2.0 HD detection kit. Therefore, this investigation starts from the need to analytically validate the new mRNA ISH assay and, possibly, to refine the current algorithm for HPV detection in oropharyngeal squamous cell carcinoma (OSCC) with the final goal to apply it to daily laboratory practice. The study was based on HPV status and on generated data, interpreted by a scoring algorithm. The results highlighted that the compared RNAscope HPV tests had a good level of interchangeability and enabled to identify OSCC that are truly driven by high risk-HPV infection. This was also supported by 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 material for HPV E6/E7 mRNA real time RT-PCR was available. Furthermore, the algorithm that associates p16 immunohistochemistry (IHC) with the identification of HPV mRNA by RNAscope was more effective than the one that associated p16 IHC with the identification of HPV DNA by ISH.



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