卵巢癌分子分析的最新进展揭示了许多有前途的肿瘤特异性生物标志物，包括蛋白质、DNA 突变和甲基化；然而，通过低成本方法以令人满意的高灵敏度和特异性可靠地检测此类改变仍然具有挑战性，特别是在早期疾病中。在这里，我们推出了 PapDREAM，这是一种新方法，能够从常规采集的宫颈巴氏标本中检测罕见的卵巢癌特异性 DNA 甲基化畸变。 PapDREAM 方法采用微流体平台，执行高度并行化的数字高分辨率熔解，以单个 CpG 位点分辨率或接近单个 CpG 位点的分辨率（< 1/10）逐个分析位点特异性 DNA 甲基化模式。下一代测序技术的成本。我们通过使用来自 43 名女性队列（包括 18 名卵巢癌患者和 25 名无癌对照）的巴氏涂片样本中的 DNA 评估一组甲基化生物标志物位点中 DNA 甲基化的分子间异质性，证明了该平台的可行性。 PapDREAM 利用位点特异性甲基化异质性的系统多维生物信息分析来改进基于巴氏样本的卵巢癌检测，在检测卵巢癌病例时，其临床敏感性为 50%，特异性为 99%，受试者工作曲线下面积为0.90。然后，我们建立了一个逻辑回归模型，可用于识别高风险患者，以便进行后续的临床随访和监测。这项研究的结果支持 PapDREAM 作为一种简单、低成本的筛查方法的实用性，并有可能与现有的临床工作流程相结合，以实现卵巢癌的早期检测。要点：我们提出了一个微流体平台，用于检测和分析巴氏标本中稀有的、异质的甲基化 DNA，以检测卵巢癌。该平台以适当较低的成本（约 25 美元）实现了常规筛查应用的高灵敏度（分数 <0.00005%）。此外，它还提供逐个分子的定量分析，以方便进一步研究异质甲基化对癌症发展的影响。© 2024 作者。约翰·威利出版的《临床与转化医学》

Recent advances in molecular analyses of ovarian cancer have revealed a wealth of promising tumour-specific biomarkers, including protein, DNA mutations and methylation; however, reliably detecting such alterations at satisfactorily high sensitivity and specificity through low-cost methods remains challenging, especially in early-stage diseases. Here we present PapDREAM, a new approach that enables detection of rare, ovarian-cancer-specific aberrations of DNA methylation from routinely-collected cervical Pap specimens. The PapDREAM approach employs a microfluidic platform that performs highly parallelized digital high-resolution melt to analyze locus-specific DNA methylation patterns on a molecule-by-molecule basis at or near single CpG-site resolution at a fraction (< 1/10th) of the cost of next-generation sequencing techniques. We demonstrate the feasibility of the platform by assessing intermolecular heterogeneity of DNA methylation in a panel of methylation biomarker loci using DNA derived from Pap specimens obtained from a cohort of 43 women, including 18 cases with ovarian cancer and 25 cancer-free controls. PapDREAM leverages systematic multidimensional bioinformatic analyses of locus-specific methylation heterogeneity to improve upon Pap-specimen-based detection of ovarian cancer, demonstrating a clinical sensitivity of 50% at 99% specificity in detecting ovarian cancer cases with an area under the receiver operator curve of 0.90. We then establish a logistic regression model that could be used to identify high-risk patients for subsequent clinical follow-up and monitoring. The results of this study support the utility of PapDREAM as a simple, low-cost screening method with the potential to integrate with existing clinical workflows for early detection of ovarian cancer. KEY POINTS: We present a microfluidic platform for detection and analysis of rare, heterogeneously methylated DNA within Pap specimens towards detection of ovarian cancer. The platform achieves high sensitivity (fractions <0.00005%) at a suitably low cost (∼$25) for routine screening applications. Furthermore, it provides molecule-by-molecule quantitative analysis to facilitate further study on the effect of heterogeneous methylation on cancer development.© 2024 The Author(s). Clinical and Translational Medicine published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics.