title: Diagnosis of HPV driven oropharyngeal cancers: Comparing p16 based algorithms with the RNAscope HPV-test.
abstract: Accurate identification of HPV-driven oropharyngeal cancer (OPC) is a major issue and none of the current diagnostic approaches is ideal. An in situ hybridization (ISH) assay that detects high-risk HPV E6/E7 mRNA, called the RNAscope HPV-test, has been recently developed. Studies have suggested that this assay may become a standard to define HPV-status.
  
  
  
  To further assess this test, we compared its performance against the strategies that are used in routine clinical practice: p16 immunohistochemistry (IHC) as a single test and algorithms combining p16-IHC with HPV-DNA identification by PCR (algorithm-1) or ISH (algorithm-2).
  
  
  
  105 OPC specimens were analyzed. The prevalence of HPV-positive samples varied considerably: 67% for p16-IHC, 54% for algorithm-1, 61% for algorithm-2 and 59% for the RNAscope HPV-test. Discrepancies between the RNAscope HPV-test and p16-IHC, algorithm-1 and 2 were noted in respectively 13.3%, 13.1%, and 8.6%. The 4 diagnostic strategies were able to identify 2 groups with different prognosis according to HPV-status, as expected. However, the greater survival differential was observed with the RNAscope HPV-test [HR: 0.19, 95% confidence interval (CI), 0.07-0.51, p=0.001] closely followed by algorithm-1 (HR: 0.23, 95% CI, 0.08-0.66, p=0.006) and algorithm-2 (HR: 0.26, 95% CI, 0.1-0.65, p=0.004). In contrast, a weaker association was found when p16-IHC was used as a single test (HR: 0.33, 95% CI, 0.13-0.81, p=0.02).
  
  
  
  Our findings suggest that the RNAscope HPV-test and p16-based algorithms perform better that p16 alone to identify OPC that are truly driven by HPV-infection. The RNAscope HPV-test has the advantage of being a single test.
authors.affiliation: Department of Otolaryngology - Head and Neck Surgery, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France. Electronic address: haitham.mirghani@gustaveroussy.fr.
authors.forename: Haïtham
authors.initials: H
authors.lastname: Mirghani
authors.affiliation: Department of Biopathology, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France.
authors.forename: Odile
authors.initials: O
authors.lastname: Casiraghi
authors.affiliation: Department of Otolaryngology - Head and Neck Surgery, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France.
authors.forename: Joanne
authors.initials: J
authors.lastname: Guerlain
authors.affiliation: Department of Otolaryngology, Peterborough City Hospital and Addenbrooke's Hospital, Cambridge, UK.
authors.forename: Furrat
authors.initials: F
authors.lastname: Amen
authors.affiliation: Advanced Cell Diagnostics, 3960 Point Eden Way, Hayward, CA 94545, USA.
authors.forename: Ming-Xiao
authors.initials: MX
authors.lastname: He
authors.affiliation: Advanced Cell Diagnostics, 3960 Point Eden Way, Hayward, CA 94545, USA.
authors.forename: Xiao-Jun
authors.initials: XJ
authors.lastname: Ma
authors.affiliation: Advanced Cell Diagnostics, 3960 Point Eden Way, Hayward, CA 94545, USA.
authors.forename: Yuling
authors.initials: Y
authors.lastname: Luo
authors.affiliation: INSERM UMR-S 903, SFR CAP-Santé FED 4231, Université de Reims Champagne-Ardenne, F-51100 Reims, France.
authors.forename: Céline
authors.initials: C
authors.lastname: Mourareau
authors.affiliation: Biobank, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France.
authors.forename: Françoise
authors.initials: F
authors.lastname: Drusch
authors.affiliation: Department of Biopathology, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France.
authors.forename: Aïcha Ben
authors.initials: AB
authors.lastname: Lakdhar
authors.affiliation: Department of Otolaryngology - Head and Neck Surgery, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France.
authors.forename: Antoine
authors.initials: A
authors.lastname: Melkane
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, Hôpital Tenon, Assistance Publique-Hôpitaux de Paris, France.
authors.forename: Lacau
authors.initials: L
authors.lastname: St Guily
authors.affiliation: Department of Pathology, Hôpital Européen Georges Pompidou, Assistance Publique-Hôpitaux de Paris, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Badoual
authors.affiliation: Department of Biopathology, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France; Biobank, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France; Laboratory of Translational Research, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France.
authors.forename: Jean Yves
authors.initials: JY
authors.lastname: Scoazec
authors.affiliation: Department of Biostatistics and Epidemiology, Gustave Roussy Cancer Campus and University Paris-Sud, Villejuif, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Borget
authors.affiliation: Department of Biostatistics and Epidemiology, Gustave Roussy Cancer Campus and University Paris-Sud, Villejuif, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Aupérin
authors.affiliation: INSERM UMR-S 903, SFR CAP-Santé FED 4231, Université de Reims Champagne-Ardenne, F-51100 Reims, France.
authors.forename: Veronique
authors.initials: V
authors.lastname: Dalstein
authors.affiliation: Department of Biopathology, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France; Laboratory of Translational Research, Gustave Roussy Cancer Campus, 114 rue Edouard Vaillant, Villejuif, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Vielh
mesh_terms: D000328:Adult; D000465:Algorithms; D052685:Alphapapillomavirus / Q000302:isolation & purification*; D002294:Carcinoma, Squamous Cell / Q000175:diagnosis* / Q000821:virology; D005260:Female; D006801:Humans; D017403:In Situ Hybridization; D008297:Male; D008875:Middle Aged; D009959:Oropharyngeal Neoplasms / Q000175:diagnosis* / Q000821:virology
keywords: E6/E7 mRNA/ transcripts; Head and neck cancer; Human papillomavirus 16; In situ hybridization; Oropharyngeal; Oropharynx; P16 immunostaining; RNAscope HPV-test®