title: p16/Ki-67 dual staining in cervico-vaginal cytology: correlation with histology, Human Papillomavirus detection and genotyping in women undergoing colposcopy.
abstract: OBJECTIVE
  To evaluate the CINtec PLUS assay (mtm laboratories), a new immunocytochemical method for the simultaneous detection of p16(INK4a) and Ki-67, in liquid-based cervico-vaginal cytology, investigating the association of the dual staining with HPV infection and genotyping as well as cytological and histological abnormalities.
  
  
  METHODS
  140 women with a cervico-vaginal sample obtained immediately before the colposcopy were enrolled. This cytological sample was used for HPV testing with the Linear Array HPV Genotyping Test, the dual staining with the CINtec PLUS kit and the morphology assessment.
  
  
  RESULTS
  Cytology results were 38 NILM, 16 ASC-US, 32L-SIL, 54H-SIL or worse. 113 patients also had a colposcopy-guided biopsy, classified as 14 negative, 35 CIN1, 24 CIN2, 37 CIN3, 3 invasive SCC. A strong association between p16/Ki-67 and HR-HPV infection was found (COR=6.86, 95% CI: 1.84-31.14). Importantly, the association between p16/Ki-67 positivity and HPV16 and/or 18 infection was 2-fold stronger compared to that with the infection by other HR-HPV types (COR=9.92, 95% CI: 2.39-47.77 vs COR=4.20, 95% CI: 0.99-20.87). In addition, p16/Ki-67 positivity rate significantly increased with the severity of the cytological and histological abnormalities (p<0.05 in both cases). p16/Ki-67 positivity resulted strongly associated with a CIN2+ diagnosis (COR=10.86 95% CI: 4.16-29.12).
  
  
  CONCLUSIONS
  This preliminary study evidenced that p16/Ki-67 immunostaining might have a relevant clinical role, since the dual staining was significantly associated with HR-HPV infection, particularly with HPV 16 and 18, and the increasing grade of the cervical lesions, the positivity for this biomarker being strongly related to the presence of a CIN2+ lesion.
authors.affiliation: STI Unit, San Gallicano Dermatological Institute, via Elio Chianesi 53, 00144, Rome, Italy.
authors.forename: Maria Gabriella
authors.initials: MG
authors.lastname: Donà
authors.forename: Amina
authors.initials: A
authors.lastname: Vocaturo
authors.forename: Massimo
authors.initials: M
authors.lastname: Giuliani
authors.forename: Livia
authors.initials: L
authors.lastname: Ronchetti
authors.forename: Francesca
authors.initials: F
authors.lastname: Rollo
authors.forename: Edoardo
authors.initials: E
authors.lastname: Pescarmona
authors.forename: Mariantonia
authors.initials: M
authors.lastname: Carosi
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Vocaturo
authors.forename: Maria
authors.initials: M
authors.lastname: Benevolo
mesh_terms: D000328:Adult; D000368:Aged; D002584:Cervix Uteri / Q000166:cytology* / Q000473:pathology / Q000821:virology; D003127:Colposcopy / Q000379:methods; D019941:Cyclin-Dependent Kinase Inhibitor p16; D003581:Cytodiagnosis / Q000706:statistics & numerical data; D005260:Female; D005838:Genotype; D006801:Humans; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry; D019394:Ki-67 Antigen / Q000737:chemistry / Q000378:metabolism*; D008875:Middle Aged; D009363:Neoplasm Proteins / Q000737:chemistry / Q000378:metabolism*; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000473:pathology / Q000821:virology; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis* / Q000378:metabolism / Q000473:pathology / Q000821:virology; D014621:Vagina / Q000166:cytology* / Q000473:pathology / Q000821:virology; D014626:Vaginal Smears / Q000379:methods; D055815:Young Adult
chemical_list: C000614131:CDKN2A protein, human; D019941:Cyclin-Dependent Kinase Inhibitor p16; D019394:Ki-67 Antigen; D009363:Neoplasm Proteins