Background More than 95% of cervical cancers and their precancerous lesions are caused by human papillomavirus (HPV). Cell-free (cf) HPV DNA detection in blood samples may serve as a monitoring tool for cervical cancer. Methods In our methodological study, an HPV panel for simultaneous detection of 24 types using mass spectrometry-based analysis was developed for liquid biopsy approaches and tested on HPV positive cell lines, plasmid controls, and cervical high-grade squamous intraepithelial lesions (HSIL) in positive smear samples (n = 52). It was validated in cfDNA blood samples (n = 40) of cervical cancer patients. Results The HPV panel showed proficient results in cell lines and viral plasmids with a limit of detection of 1 IU (international units)/µL for HPV16/18 and 10GE/µL for HPV11/31/33/39/45/51/52/58/59 and a specificity of 100% for the tested HPV types. In cervical smear samples, HPV DNA was detected with a sensitivity of 98.14%. The overall agreement between the new HPV panel and clinical records was 97.2% (κ = 0.84). In cervical cancer cfDNA, 26/40 (65.0%) tested positive for any HPV type, with most infections due to hrHPV (24/26). HPV positive samples were found in all FIGO stages, with the highest positivity ratio in FIGO III and IV. Even the lowest stage, FIGO I, had 12/23 (52.2%) patients with a positive HPV plasma status. Conclusions This proof-of-concept paper shows that the described assay produces reliable results for detecting HPV types in a multiplex mass spectrometry-based assay in cervical smear and cfDNA with high specificity and sensitivity in both cohorts. The assay shows potential for liquid biopsy-based applications in monitoring cervical cancer progression.

中文翻译：

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宫颈肿瘤液体活检中多种 HPV 类型的检测

背景 95%以上的宫颈癌及其癌前病变是由人乳头瘤病毒（HPV）引起的。血液样本中的无细胞 (cf) HPV DNA 检测可作为宫颈癌的监测工具。方法 在我们的方法学研究中，针对液体活检方法开发了一种使用基于质谱分析同时检测 24 种类型的 HPV 检测组，并在 HPV 阳性细胞系、质粒对照和宫颈高级鳞状上皮内病变 (HSIL) 中进行了测试。阳性涂片样本 (n = 52)。它在宫颈癌患者的 cfDNA 血液样本 (n = 40) 中得到了验证。结果 HPV panel 在细胞系和病毒质粒中显示出良好的结果，HPV16/18 的检测限为 1 IU（国际单位）/μL，HPV11/31/33/39/45/51/52/ 的检测限为 10GE/μL。 58/59，所测试 HPV 类型的特异性为 100%。在宫颈涂片样本中，HPV DNA 的检测灵敏度为 98.14%。新的 HPV 检测结果与临床记录之间的总体一致性为 97.2% (κ = 0.84)。在宫颈癌 cfDNA 中，26/40 (65.0%) 的任何 HPV 类型检测呈阳性，其中大多数感染由 hrHPV (24/26) 引起。所有FIGO阶段均发现HPV阳性样本，其中FIGO III和IV阶段阳性率最高。即使是最低阶段，FIGO I，也有 12/23 (52.2%) 的患者 HPV 血浆状态呈阳性。结论 这篇概念验证论文表明，在基于宫颈涂片和 cfDNA 的多重质谱检测中，所描述的检测方法在检测 HPV 类型方面产生了可靠的结果，在两个队列中具有高特异性和敏感性。该测定显示了基于液体活检的应用在监测宫颈癌进展方面的潜力。