准确评估个体宫颈病变风险，对精确指导筛查和治疗至关重要。HPV DNA检测灵敏度高，是广泛应用的初筛方法，但其有限的特异度使阳性人群分流成为必要。研究显示HPV E6/E7癌蛋白等能一定程度分流DNA阳性和有进展风险的对象，但其检测性质不同，难以同时实现，耗时长且成本高。靶向性测序检测连续甲基化是基于二代测序的定性检测，能同时检测多个位点连续甲基化状况，并整合病毒及宿主的信息，灵敏度高且不受采样偏倚干扰。本研究拟利用该平台完成多靶标检测 “HPV DNA”+“HPV E6/E7癌蛋白上下游通路分子连续性甲基化”，在完成初筛的同时实现分流。首先建立针对多重差异甲基化目的区段的富集和测序平台，引入机器学习，结合前期检测结果，构建多靶点、综合初筛和分流功能的靶向测序技术；随后在三千人宫颈癌筛查队列中，经3年随访，以传统筛查方法为参照评价其在宫颈癌筛查中的应用价值，建立针对不同风险人群的防治策略。

Accurate risk assessment of individual cervical lesion is crucial for precise screening and treatment. Detection of HPV DNA has been widely used for primary screening owing to its high sensitivity. However, the test has limited specificity, therefore demanding for additional triage assays for HPV positive subjects. It has been shown that HPV oncoprotein E6/E7 can potentially help identify cancer risk in HPV positive patients. Unfortunately, it is not feasible to perform all tests in a single reaction, rendering clinical triage time-consuming and costly. Targeted DNA methylation has recently shown as a quality test with great potential in clinical practice. Based on NextGen sequencing, it can detects continuous methylation at multiple CpG sites simultaneously, integrates virus and host information, and has high detection sensitivity and little or none sampling bias. This study will perform targeted sequencing analysis of multiple loci of interest (HPV DNA plus continuous methylation of genes involved in HPV E6/E7 oncogenic pathway) to achieve primary screening and patient triage in one test. Firstly, we aim to establish an innovative platform for multiplex enrichment of differentially methylated segments for deep sequencing. Taking advantage of a well-annotated cross-sectional patient, machine learning will be used to deliver a multi-target DNA methylation test for primary cervical cancer screening and patient triage. In the second stage, the clinical utility of the aforementioned test will be validated using an independent cervical cancer screening cohort of 3,000 subjects. The test performance will be evaluated by comparing with the traditional screening method through three-year follow-up. As the result, we expect to establish a novel screening strategy for the prevention and treatment of HPV patients in different risk groups.