Ultra-sensitive urine DNA methylation test enables early and accurate detection of bladder cancer

作者：Zhichao Tong, Tianqi Yu, Zhilong Li, Fengming Zhang, Shi Qy, Jiaxi Peng, Yu Zhao, Yaowei Li, Zhihui Xiu, Zhishuai Zhang, Benchi Li, Weijia Jiang, Hongjian Song, Zongzheng Yang, Hui Li, Pengyu Guo, Dayong Hou, Zhihao Yin, Liu Z, Qi Liu, Yi-Xiang Wang, P Zhang, Peng Dai, Yuying Wang, Di Wang, Hang Su, Ziqi Wang · 发表于：Epigenomics · 年份：2026 · DOI：10.1080/17501911.2026.2688972 · 研究领域：Bladder and Urothelial Cancer Treatments、Epigenetics and DNA Methylation、Urinary and Genital Oncology Studies

Background Early and accurate detection of bladder cancer (BCa), particularly low-grade and early-stage disease, remains challenging because current diagnostic methods are either invasive or insufficiently sensitive. We aimed to develop a noninvasive urine DNA methylation assay for BCa detection and postoperative monitoring.Research design and methods Integrative methylation profiling across institutional and public cohorts identified PENK and NKPD1 as urothelial carcinoma-associated hypermethylation markers. A bisulfite-free methylation-sensitive restriction enzyme quantitative polymerase chain reaction (MSRE-qPCR) assay was developed using urine sediment DNA. Diagnostic performance was evaluated in a case-control cohort of 606 urine samples, including 340 pathologically confirmed BCa cases.Results The dual-marker assay achieved areas under the curve (AUCs) of 92.52% and 93.00% in the training and validation cohorts, respectively. Sensitivity reached 87.74%, with specificity of 92.31% against benign urological diseases and 83.05% against non-urothelial malignancies. The assay outperformed urine cytology in low-grade tumors and provided complementary diagnostic value when combined with cytology. Postoperative methylation signals significantly decreased after tumor resection (p < 0.001).Conclusions This bisulfite-free urine MSRE-qPCR assay provides a simple and scalable approach for noninvasive BCa detection and surveillance, with strong performance in early-stage and low-grad...