Bulfoni Michela, Tel Alessandro, Krpan Beatrice, Marcon Barbara, De Martino Maria, Vesca Giorgia, Lombardo Cinzia, Marzinotto Stefania, Nencioni Emanuele, Sembronio Salvatore, Pipan Corrado, Robiony Massimo, Curcio Francesco
Department of Medicine, University of Udine, Udine, Italy.
Department of Head and Neck and Neuroscience, Academic Hospital of Udine, Clinic of Maxillofacial Surgery, Udine, Italy.
Head Neck. 2025 Aug 5. doi: 10.1002/hed.70003.
Human papilloma virus (HPV) is a DNA virus capable of infecting mucous membranes. In most cases, the infection is cleared by the immune system, but a prolonged exposure to HPV can progress to cancer. 40%-60% of head and neck squamous cell carcinomas (HNSCC) are linked to HPV, which is considered a risk factor especially among young people in industrialized countries. HNSCC are not identified early due to their slow growth and their locations not being easy to see. Despite the prognostic value of HPV, the use of HPV-DNA as a diagnostic marker is not fully developed. HPV-DNA can be detected in saliva specimens of patients with HPV-driven cancers. Considering this, we employed saliva samples to optimize a non-invasive RT-PCR assay for HPV infection and prove the hypothesis that the presence of HPV DNA represents a risk factor for Oral Cavity Squamous Cell Carcinoma (OCSCC). The potential of this work is to highlight how an HPV screening program based on salivary testing can be useful for early cancer detection and patient monitoring.
In this retrospective study, 127 patients and 93 control samples were tested for the presence of HPV in self-collected saliva specimens. Viral DNA was extracted from saliva using an automated instrument. A multiplex RT-PCR was employed for the detection of the (28 most frequent HPV Types). Patients' demographics were collected in a clinical database. Statistics were performed by STATA16, and significance was set at p < 0.05.
Most patients had a diagnosis of OCSCC (49.1%), 38%-9% of which involved the tongue. 9.6% resulted positive for HPV DNA in saliva, specifically for high-risk subtypes (42.9% type 58; 28.6% types 45, 59, 39 and 9.6% type 16, 18). HPV+patients were compared to those who were found negative. HPV infection was related to the TNM stage, especially with pT2N1 (p ≤ 0.002) and with primary vs. relapsed tumors (p = 0.004). Independent of site, the assay reached a sensitivity of 91.7%, a specificity of 100%, and an agreement of 98.5%, compared to the oropharyngeal swab (Cohen's Kappa = 0.947; n = 65). Test PPV was 100% (95% CI 71.5-100), while NPV was 98.1% (95% CI 90.1-100).
Our findings indicate that salivary HPV testing is a non-invasive and convenient test that could become part of routine clinical management for HPV infection of the oral cavity. This test represents an ideal mode of screening of asymptomatic individuals and a long-term monitoring tool for HPV-driven cancer patients.
人乳头瘤病毒（HPV）是一种能够感染黏膜的DNA病毒。在大多数情况下，感染会被免疫系统清除，但长期接触HPV可能会发展为癌症。40%-60%的头颈部鳞状细胞癌（HNSCC）与HPV有关，在工业化国家，这被认为是一个风险因素，尤其是在年轻人中。由于HNSCC生长缓慢且位置不易观察，所以难以早期发现。尽管HPV具有预后价值，但将HPV-DNA用作诊断标志物的应用尚未得到充分发展。在HPV驱动的癌症患者的唾液样本中可以检测到HPV-DNA。考虑到这一点，我们使用唾液样本优化了一种用于HPV感染的非侵入性逆转录聚合酶链反应（RT-PCR）检测方法，并证明了HPV DNA的存在是口腔鳞状细胞癌（OCSCC）的一个风险因素这一假设。这项工作的潜力在于突出基于唾液检测的HPV筛查计划如何有助于早期癌症检测和患者监测。
在这项回顾性研究中，对127例患者和93份对照样本的自行采集的唾液样本进行了HPV检测。使用自动化仪器从唾液中提取病毒DNA。采用多重RT-PCR检测（28种最常见的HPV类型）。患者的人口统计学数据收集在临床数据库中。使用STATA16进行统计分析，显著性设定为p < 0.05。
大多数患者被诊断为OCSCC（49.1%），其中38%-9%累及舌头。9.6%的患者唾液中的HPV DNA检测呈阳性，具体为高危亚型（58型占42.9%；45、59、39型占28.6%，16、18型占9.6%）。将HPV阳性患者与检测为阴性的患者进行比较。HPV感染与TNM分期有关，尤其是与pT2N1（p≤0.002）以及原发性肿瘤与复发性肿瘤有关（p = 0.004）。与口咽拭子相比（Cohen's Kappa = 0.947；n = 65），该检测方法无论在哪个部位，灵敏度均达到91.7%，特异性为100%，一致性为98.5%。检测的阳性预测值（PPV）为100%（95%可信区间71.5 - 100），阴性预测值（NPV）为98.1%（95%可信区间90.1 - 100）。
我们的研究结果表明，唾液HPV检测是一种非侵入性且便捷的检测方法，可成为口腔HPV感染常规临床管理的一部分。该检测方法是无症状个体筛查的理想方式，也是HPV驱动的癌症患者的长期监测工具。
