Department of Pathology, Microbiology, and Immunology and Vanderbilt University Medical Center, Nashville, TN, US.
Department of Otolaryngology-Head and Neck Surgery, Vanderbilt University Medical Center, Nashville, TN, US.
Am J Clin Pathol. 2024 Jun 3;161(6):570-578. doi: 10.1093/ajcp/aqad185.
In this feasibility study, we explored the combined use of circulating tumor human papillomavirus (HPV) DNA (ctHPVDNA) and HPV serology as diagnostic tests for HPV-associated oropharyngeal squamous cell carcinoma (OPSCC).
Among patients with research-banked serum or plasma at diagnosis, IgG antibodies to oncoproteins from HPV types 16, 18, 31, 33, 35, 45, 52, and 58 were detected with multiplex serology. Positivity for HPV 16 was defined based on detection of combinations of anti-E6, E1, E2, and E7 and for other high-risk types on detection of anti-E6 and anti-E7. Circulating tumor HPV DNA was detected by custom digital droplet polymerase chain reaction (ddPCR) assays for HPV types 16, 18, 33, 35, and 45. p16 immunohistochemistry and high-risk HPV RNA in situ hybridization (ISH) using a cocktail of 18 high-risk HPV types were performed on tissue.
Of 75 patients, 67 (89.3%) were HPV-associated (p16 and HPV RNA ISH positive) and 8 (10.7%) were HPV-independent. All 8 HPV-independent patients were seronegative and negative for ctHPVDNA (100% specificity). Serology was positive in 53 (79.1%) of 67 HPV-associated patients, while ddPCR was positive for ctHPVDNA in 59 (88.6%) of 67 HPV-associated patients. Requiring both tests to be positive resulted in a sensitivity of 50 (74.6%) of 67 while combining assays (either positive) improved sensitivity to 62 (92.6%) of 67.
Compared to HPV RNA ISH, HPV serology and ctHPVDNA are sensitive and highly specific biomarkers for HPV-associated OPSCC at the time of presentation.
在这项可行性研究中，我们探讨了联合使用循环肿瘤人乳头瘤病毒（HPV）DNA（ctHPV DNA）和 HPV 血清学作为 HPV 相关口咽鳞状细胞癌（OPSCC）的诊断检测方法。
在有研究支持的血清或血浆样本的患者中，使用多重血清学方法检测 HPV 型 16、18、31、33、35、45、52 和 58 的致癌蛋白的 IgG 抗体。HPV 16 的阳性定义为检测到抗 E6、E1、E2 和 E7 的组合，而对于其他高危型则检测到抗 E6 和抗 E7。通过定制的数字液滴聚合酶链反应（ddPCR）检测 HPV 型 16、18、33、35 和 45 的循环肿瘤 HPV DNA。对组织进行 p16 免疫组化和使用 18 种高危型 HPV 混合物的高危 HPV RNA 原位杂交（ISH）。
在 75 例患者中，67 例（89.3%）为 HPV 相关（p16 和 HPV RNA ISH 阳性），8 例（10.7%）为 HPV 不相关。所有 8 例 HPV 不相关的患者均为血清学阴性且 ctHPV DNA 阴性（特异性 100%）。67 例 HPV 相关患者中，53 例（79.1%）血清学阳性，67 例 HPV 相关患者中，59 例（88.6%） ddPCR 检测到 ctHPV DNA 阳性。要求两种检测均为阳性时，敏感性为 67 例中的 50 例（74.6%），而联合检测（任一种阳性）则将敏感性提高到 67 例中的 62 例（92.6%）。
与 HPV RNA ISH 相比，HPV 血清学和 ctHPV DNA 是在出现时诊断 HPV 相关 OPSCC 的敏感且高度特异的生物标志物。
