ArticleJCO precision oncology2026
Molecular Etiology and Clinical Significance of Discordant Human Papillomavirus Testing in p16+ Oropharyngeal Cancers.
Article in JCO precision oncology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
purposeRoutine confirmatory testing of p16+ oropharyngeal cancers (OPCs) for human papillomavirus (HPV) mRNA is nonstandard despite evidence that negative confirmatory tests are common and may indicate poor prognosis. This study aimed to evaluate the utility of confirmatory testing by assessing the etiologies and prognostic significance of negative HPV mRNA tests in a surgically treated cohort of p16+ OPCs enriched for recurrence-prone tumors.
methodsFifty tumors that later recurred (cases) and 50 cured tumors (controls) were matched for key clinical features. Patients were predominantly White males with a median age of 62 years. Confirmatory testing was performed with a clinically used E6/E7 RNA in situ hybridization assay (RNAScope). RNA sequencing (RNAseq) was used to define true HPV positivity and profile HPV mRNA levels. HPV genotypes and integration were evaluated by sequencing HPV DNA and detecting HPV-host fusion mRNAs, respectively.
resultsPositive p16 was 100% specific for high-risk HPV mRNA expression. Content of HPV types, lineages, and sublineages did not differ significantly between cases and controls. Cases contained more frequent HPV integration based on increased HPV-host mRNA fusions (
conclusionHPV RNAScope appears unlikely to detect HPV status misclassification or be prognostic in typical p16+ OPCs treated surgically in the United States. Confirmatory RNAScope can produce misleading false negatives in this population through various mechanisms and should be used selectively rather than routinely.
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