title: Chromogenic in situ hybridization and p16/Ki67 dual staining on formalin-fixed paraffin-embedded cervical specimens: correlation with HPV-DNA test, E6/E7 mRNA test, and potential clinical applications.
abstract: Although HPV-DNA test and E6/E7 mRNA analyses remain the current standard for the confirmation of human papillomavirus (HPV) infections in cytological specimens, no universally adopted techniques exist for the detection of HPV in formalin-fixed paraffin-embedded samples. Particularly, in routine laboratories, molecular assays are still time-consuming and would require a high level of expertise. In this study, we investigated the possible use of a novel HPV tyramide-based chromogenic in situ hybridization (CISH) technology to locate HPV on tissue specimens. Then, we evaluate the potential usefulness of p16(INK4a)/Ki-67 double stain on histological samples, to identify cervical cells expressing HPV E6/E7 oncogenes. In our series, CISH showed a clear signal in 95.2% of the specimens and reached a sensitivity of 86.5%. CISH positivity always matched with HPV-DNA positivity, while 100% of cases with punctated signal joined with cervical intraepithelial neoplasia grade 2 or worse (CIN2+). p16/Ki67 immunohistochemistry gave an interpretable result in 100% of the cases. The use of dual stain significantly increased the agreement between pathologists, which reached 100%. Concordance between dual stain and E6/E7 mRNA test was 89%. In our series, both CISH and p16(INK4a)/Ki67 dual stain demonstrated high grade of performances. In particular, CISH would help to distinguish episomal from integrated HPV, in order to allow conclusions regarding the prognosis of the lesion, while p16(INK4a)/Ki67 dual stain approach would confer a high level of standardization to the diagnostic procedure.
authors.affiliation: Cytopathology Unit, Experimental and Clinical Sciences Department, "G. d'Annunzio" University of Chieti-Pescara, Via dei Vestini, 66100 Chieti, Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: Zappacosta
authors.affiliation: Surgical Pathology Unit, "SS Annunziata" Hospital, ASL2 Abruzzo, Chieti, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Colasante
authors.affiliation: Surgical Pathology Unit, "SS Annunziata" Hospital, ASL2 Abruzzo, Chieti, Italy.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Viola
authors.affiliation: Surgical Pathology Unit, "SS Annunziata" Hospital, ASL2 Abruzzo, Chieti, Italy.
authors.forename: Tommaso
authors.initials: T
authors.lastname: D'Antuono
authors.affiliation: Surgical Pathology Unit, "SS Annunziata" Hospital, ASL2 Abruzzo, Chieti, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Lattanzio
authors.affiliation: Surgical Pathology Unit, "SS Annunziata" Hospital, ASL2 Abruzzo, Chieti, Italy.
authors.forename: Serena
authors.initials: S
authors.lastname: Capanna
authors.affiliation: Cytopathology Unit, Experimental and Clinical Sciences Department, "G. d'Annunzio" University of Chieti-Pescara, Via dei Vestini, 66100 Chieti, Italy.
authors.forename: Daniela Maria Pia
authors.initials: DM
authors.lastname: Gatta
authors.affiliation: Cytopathology Unit, Experimental and Clinical Sciences Department, "G. d'Annunzio" University of Chieti-Pescara, Via dei Vestini, 66100 Chieti, Italy.
authors.forename: Sandra
authors.initials: S
authors.lastname: Rosini
mesh_terms: D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics / Q000302:isolation & purification*; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D019394:Ki-67 Antigen / Q000235:genetics* / Q000302:isolation & purification; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000302:isolation & purification*; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification; D030361:Papillomavirus Infections / Q000175:diagnosis / Q000235:genetics / Q000473:pathology / Q000821:virology; D016612:Paraffin Embedding; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology / Q000821:virology
chemical_list: D019941:Cyclin-Dependent Kinase Inhibitor p16; D004279:DNA, Viral; D019394:Ki-67 Antigen; D009856:Oncogene Proteins, Viral