Combination of p16 INK4a -Ki67 Immunocytology and HPV Polymerase Chain Reaction for the Noninvasive Analysis of HPV Involvement in Head and Neck Cancer

INK4a -Ki67 staining using a CINtec PLUS kit after the presence of tumor cells was verified in a Papanicolaou-stained slide. The same cytological suspension was used for the detection of HPV DNA by specific polymerase chain reaction (PCR). RESULTS: Tumor cells were detected in the swab material of 44 HNSCC patients corresponding to a sensitivity of 98% (44 of 45). PCR analysis revealed the presence of HPV DNA in the cytological suspension of 13 patients (13 of 65, 20%) with simultaneous p16 INK4a -Ki67 expression by the tumor cells in 11 of these HPV DNA–positive samples (11 of 13, 85%) — a staining pattern that is strongly associated with a carcinogenic HPV infection. CONCLUSIONS: A simultaneous immunocytochemical detection of p16 INK4a and Ki67 can reliably be performed on liquid-based cytological smears from HNSCC patients using a CINtec PLUS kit. In addition, the same cytological material can be used for the detection of HPV DNA by specific PCR. The combined results of both techniques enable better discrimination between latent and carcinogenic HPV infections as well as HPV-negative cases and thus can provide information on the prognosis of HNSCC patients and facilitate therapeutic decisions. Cancer (Cancer Cytopathol) 2014;000:000-000. V C 2014 American Cancer Society.