A tissue microarray was constructed using JHH OCC samples with available formalin-fixed, paraffin-embedded (FFPE) tissue blocks. Antigen retrieval was performed on deparaffinized, 4 micron sections using 10 mM citrate buffer at 92°C for 30 minutes. A mouse monoclonal antibody for MCPyV Large T-antigen (Santa Cruz Biotechnology, Dallas, TX; clone CM2B4; 1:500 dilution) was applied using a BenchMark Ultra autostainer (Ventana Medical Systems, Tucson, AZ). Signals were visualized using the UltraView polymer detection kit (Ventana). Staining was performed according to manufacturer’s instructions with a known positive case of Merkel cell carcinoma as a positive control and benign tonsillar tissue as a negative control. The presence of staining was assessed by a head and neck pathologist (LR).
Human papillomavirus (HPV) testing was performed on evaluable whole-slide specimens from all sites using p16 immunohistochemistry and HPV RNA in situ hybridization (ISH) for high-risk types as described elsewhere9 (link).
Human papillomavirus (HPV) testing was performed on evaluable whole-slide specimens from all sites using p16 immunohistochemistry and HPV RNA in situ hybridization (ISH) for high-risk types as described elsewhere9 (link).
