title: Human papillomavirus (HPV) infection in squamous cell carcinomas arising from the oropharynx: detection of HPV DNA and p16 immunohistochemistry as diagnostic and prognostic indicators--a pilot study.
abstract: OBJECTIVE
  Human papillomavirus (HPV) 16 infection is associated with oropharyngeal carcinogenesis and is likely the cause of the reported increase in disease incidence. We evaluated the prevalence of HPV infection and the reliability of different diagnostic tools using primary tumor samples from a cohort of 50 patients.
  
  
  METHODS
  Formalin-fixed paraffin-embedded (FFPE) tumor samples were collected from all 50 consecutive primary oropharyngeal SCC patients who were enrolled in the study; fresh tumor samples were available in 22 cases. NucliSENS EasyQ HPVv1 was used for RNA, and Digene Hybrid Capture-2(HC2) was used for DNA detection. p16 Expression was evaluated by immunohistochemistry in FPPE specimens.
  
  
  RESULTS
  Based on the DNA detection assay on FFPE samples, the frequency of high-risk HPV infection was 32%. The agreement rate between HPV RNA and HPV DNA detection in fresh samples was 100%. The agreement rate between p16 immunohistochemistry (IHC) and the detection of HPV DNA in the FFPE samples was fair but not excellent (κ = 0.618). HPV DNA detection was highly significant, as measured by disease-specific survival and determined using a Wilcoxon test (P=.001). p16 IHC also exhibited a prognostic value but with a lower statistical significance (P=.0475). The detection of HPV DNA, but not p16 IHC, was also significantly correlated with locoregional control (P=.0461).
  
  
  CONCLUSIONS
  Diagnostic methods based on the detection of HPV nucleic acids appear to be more reliable and objective because they do not require reading by a trained histopathologist. Furthermore, the detection of HPV DNA exhibits an improved correlation with survival, and therefore appears definitely more reliable than p16 IHC for routine use in clinical practice.
authors.affiliation: Institute of Otolaryngology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma, Italy. Electronic address: francesco.bussu.md@gmail.com.
authors.forename: Francesco
authors.initials: F
authors.lastname: Bussu
authors.affiliation: Institute of Microbiology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma, Italy.
authors.forename: Michela
authors.initials: M
authors.lastname: Sali
authors.affiliation: Institute of Otolaryngology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma, Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Gallus
authors.affiliation: Institute of Histopathology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma, Italy.
authors.forename: Gianluigi
authors.initials: G
authors.lastname: Petrone
authors.affiliation: Institute of Histopathology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma, Italy.
authors.forename: Gian Franco
authors.initials: GF
authors.lastname: Zannoni
authors.affiliation: Institute of Radiotherapy, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma, Italy.
authors.forename: Rosa
authors.initials: R
authors.lastname: Autorino
authors.affiliation: Institute of Radiotherapy, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma, Italy.
authors.forename: Nicola
authors.initials: N
authors.lastname: Dinapoli
authors.affiliation: Institute of Microbiology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma, Italy.
authors.forename: Rosaria
authors.initials: R
authors.lastname: Santangelo
authors.affiliation: Institute of Histopathology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma, Italy.
authors.forename: Valerio Gaetano
authors.initials: VG
authors.lastname: Vellone
authors.affiliation: Institute of Otolaryngology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Graziani
authors.affiliation: Institute of Radiotherapy, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Miccichè
authors.affiliation: Institute of Otolaryngology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Almadori
authors.affiliation: Institute of Otolaryngology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma, Italy.
authors.forename: Jacopo
authors.initials: J
authors.lastname: Galli
authors.affiliation: Institute of Microbiology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Delogu
authors.affiliation: Institute of Microbiology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma, Italy.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Sanguinetti
authors.affiliation: Institute of Histopathology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma, Italy.
authors.forename: Guido
authors.initials: G
authors.lastname: Rindi
authors.affiliation: Infections and Cancer Biology Group, International Agency for Research on Cancer-World Health Organization, Lyon, France.
authors.forename: Massimo
authors.initials: M
authors.lastname: Tommasino
authors.affiliation: Institute of Radiotherapy, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma, Italy.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Valentini
authors.affiliation: Institute of Otolaryngology, Università Cattolica del Sacro Cuore, Policlinico A. Gemelli, Roma, Italy.
authors.forename: Gaetano
authors.initials: G
authors.lastname: Paludetti
mesh_terms: D000368:Aged; D002294:Carcinoma, Squamous Cell / Q000473:pathology / Q000821:virology*; D004279:DNA, Viral / Q000032:analysis*; D005260:Female; D052162:Human papillomavirus 16 / Q000235:genetics*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009959:Oropharyngeal Neoplasms / Q000473:pathology / Q000821:virology*; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000453:epidemiology / Q000821:virology; D016612:Paraffin Embedding; D010865:Pilot Projects; D015995:Prevalence; D011379:Prognosis; D012367:RNA, Viral / Q000032:analysis*; D015203:Reproducibility of Results
chemical_list: D004279:DNA, Viral; D012367:RNA, Viral