August 2025—Human papillomavirus testing has become the standard of care in head and neck squamous cell carcinoma (HNSCC) because of the unique clinical features, staging, and treatment options for HPV-associated HNSCC. HPV-associated oropharyngeal squamous cell carcinoma (OPSCC) exhibits a favorable prognosis and improved response to chemoradiation compared with conventional HNSCC and non-HPV-associated forms, and reporting the HPV status is frequently part of clinical trial enrollments. Many patients with OPSCC present with enlarged level II or III cervical lymph nodes and, as a result, cervical lymph node fine-needle aspiration is often the first, and sometimes only, tissue obtained for diagnostic testing. With a growing menu of options available to test for HPV status, including polymerase chain reaction, DNA in situ hybridization, mRNA ISH, liquid-based HPV assays, and p16 immunohistochemistry, it is not always clear when and which HPV test to use, especially when the diagnosis is made on a cytology specimen.