{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T00:26:12Z","timestamp":1760315172771,"version":"build-2065373602"},"reference-count":38,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2022,2,1]],"date-time":"2022-02-01T00:00:00Z","timestamp":1643673600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["acsjournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Cancer Cytopathology"],"published-print":{"date-parts":[[2022,5]]},"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Determining human papillomavirus (HPV) status in oropharyngeal squamous cell carcinoma can have a significant impact on treatment and clinical outcomes. Because fine\u2010needle aspiration (FNA) is typically an initial diagnostic modality in a patient workup for primary or suspected metastatic disease, immunostaining for p16 on FNA material is a promising option to determine HPV tumor status, possibly avoiding biopsies or excisions. In this study, the authors investigated the possibility of using alcohol\u2010fixed smears as a reliable alternative for reporting p16 status.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Twenty HPV\u2010associated tumors and 20 non\u2010HPV\u2013associated tumors were identified using the gold\u2010standard histologic cutoff for positivity of \u226570% strong nuclear and cytoplasmic staining. Matched FNA specimens were identified for comparison staining, and a positive p16 result was rendered on a single aspirate smear using the same cutoff of \u226570%.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>On alcohol\u2010fixed cytology smears, 16 of 20 (80%) HPV\u2010associated tumors showed positive p16 staining in \u226570% tumor cells. Four cases showed lower level (30%\u201060%) nuclear and cytoplasmic staining. Nineteen of 20 (95%) non\u2010HPV\u2013associated tumors showed no or minimal p16 staining (0%\u201010%), and 1 case had a p16\u2010equivocal cytology result.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>The authors performed immunocytochemical validation for p16 using alcohol\u2010fixed smears and observed promising results, offering this technique as a potential alternative to formalin\u2010fixed tissue in the appropriate clinical context. By using a positive staining cutoff of \u226570%, this technique offers 80% sensitivity and 95% specificity for detecting HPV\u2010associated tumors. Although it was not performed in the current study, HPV\u2010specific testing on available formalin\u2010fixed, paraffin\u2010embedded tissue should be considered in cases with equivocal or negative p16 staining.<\/jats:p><\/jats:sec>","DOI":"10.1002\/cncy.22557","type":"journal-article","created":{"date-parts":[[2022,2,1]],"date-time":"2022-02-01T09:09:07Z","timestamp":1643706547000},"page":"381-391","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Validation of p16 immunocytochemical staining on alcohol\u2010fixed aspirate smears: A single\u2010institutional study with review of the literature"],"prefix":"10.1002","volume":"130","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4873-8018","authenticated-orcid":false,"given":"Katie","family":"Doonan","sequence":"first","affiliation":[{"name":"Department of Pathology and Laboratory Medicine University of Vermont Medical Center Burlington 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