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

The aim of this study is to compare 2 in situ hybridization (ISH) detection methods for human papilloma virus (HPV) 16 E6/E7 mRNA, that is, 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. The...

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Bibliographic Details
Published in:	Human pathology 2018-04, Vol.74, p.32-42
Main Authors:	Volpi, Chiara C., Ciniselli, Chiara M., Gualeni, Ambra V., Plebani, Maddalena, Alfieri, Salvatore, Verderio, Paolo, Locati, Laura, Perrone, Federica, Quattrone, Pasquale, Carbone, Antonino, Pilotti, Silvana, Gloghini, Annunziata
Format:	Article
Language:	eng
Subjects:	Algorithms Automation Deoxyribonucleic acid DNA HPV 16 HPV E6/E7 mRNA ISH HR-HPV DNA ISH Human papillomavirus Hybridization Infections Oropharyngeal squamous cell carcinoma p16 Working groups
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Summary:	The aim of this study is to compare 2 in situ hybridization (ISH) detection methods for human papilloma virus (HPV) 16 E6/E7 mRNA, that is, 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 with the final goal of applying 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 oropharyngeal squamous cell carcinoma 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-transcription polymerase chain reaction in a fraction of cases where material for HPV E6/E7 mRNA real-time reverse-transcription polymerase chain reaction was available. Furthermore, the algorithm that associates p16 immunohistochemistry with the identification of HPV mRNA by RNAscope was more effective than the one that associated p16 immunohistochemistry with the identification of HPV DNA by ISH. •This investigation starts from the need to compare 2 HPV mRNA ISH assays.•The study also aimed at refining the current algorithm for HPV detection in OSCC.•The compared RNAscope HPV tests (2.0 versus 2.5) had a good level of interchangeability.•The algorithm associating p16 IHC with HPV mRNA ISH was the most effective.•mRNA ISH assays are able to identify OSCCs that are truly driven by HR-HPV infection.
ISSN:	0046-8177 1532-8392