Adequacy of FNA specimens for HPV molecular testing in head and neck SCC

Adequacy of fine-needle aspiration specimens for human papillomavirus infection molecular testing in head and neck squamous cell carcinoma

Ricardo R Lastra, Michelle R Pramick, Megan O Nakashima, Gregory S Weinstein, Kathleen T Montone, Virginia A LiVolsi, Zubair W Baloch

CytoJournal 2013 10(1):21-21


Head and neck squamous cell carcinoma (HNSCC) is often associated with human papillomavirus (HPV) infection. Positive HPV status has been associated with increased response to treatment and improved prognosis in terms of recurrence-free and overall survival. In certain instances, diagnosis is performed through fine-needle aspiration of lymph nodes with metastatic carcinoma, often demonstrating extensive tumor necrosis. We evaluated the effect of tumor necrosis on deoxyribonucleic acid (DNA) adequacy for HPV molecular testing.

Materials and Methods:

Retrospective review of the pathology files at our institution identified cases of squamous cell carcinoma (SCC) diagnosed by fine-needle aspiration (FNA) on which HPV DNA molecular testing was performed. The cases were classified according to percent tumor necrosis into three categories (<10% necrosis, 10-70% necrosis and >70% necrosis) and the percentage of cases with adequate HPV DNA for molecular testing in each of the categories was compared. When available, p16 immunohistochemistry performed on the cases was compared with HPV status by molecular testing.


A total of 70 cases from 67 patients were included in the study. Adequate DNA for molecular HPV testing was obtained from samples of 47 cases (67%) while samples from 23 cases (33%) were inadequate for molecular testing. Of the adequate samples, 36 (77%) were positive and 11 (23%) were negative for high-risk HPV. Adequate DNA for testing was obtained in 22 out of 33 cases showing no necrosis (67%), 10 out of 16 cases showing partial necrosis (63%) and in 13 out of 17 cases showing extensive necrosis (76%).


Our study found that HPV molecular testing is not influenced by percent tumor necrosis or method by which FNA was performed. We believe that a portion of the FNA specimen obtained from head and neck lesions diagnosed as SCC during the rapid on-site evaluation should be sent for HPV DNA testing, independent of the amount of tumor necrosis, thus guaranteeing availability of specimen for HPV testing.
(via Articles : CytoJournal as on October 30, 2013)

The authors offer a useful practical article demonstrating the effect of necrosis on the adequacy of FNA samples for molecular HPV testing in head and neck SCC.  The incidence of HNSCC is on the rise, and HPV related HNSCC tends to occur in younger patients.

” … [the] significant difference in the outcome between HPV-positive and HPV-negative HNSCC has made it a common practice to evaluate these lesions for the presence of underlying HPV infection, either by the use of surrogate markers such as p16 immunohistochemistry or by HPV DNA detection by molecular studies.”

FNA samples offer an opportunity for HPV testing when the samples are necrotic, and usual p16 stains on cell blocks cannot be performed. As cytopathology becomes more focused on personalized health care, identifying optimal samples for molecular testing on cytologic samples is essential.

The Populist Cytologist


About Jana Sullinger

GYN Pathologist and Cytopathologist. Special interest in quality assessment and use of online social media in sharing educational information about cytology and quality.


  1. […] Adequacy of FNA specimens for HPV molecular testing in head and neck SCC The Populist Cytologist™ […]

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