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Ultra-sensitive urine DNA methylation test enables early and accurate detection of bladder cancer
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DOI：10.1080/17501911.2026.2688972.png)
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Abstract
En 中文
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.
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.
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).
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-grade disease. Further prospective multicenter validation is warranted.
Keywords:
Urinary bladder neoplasms
DNA methylation
urine biomarkers
early detection of cancer
noninvasive diagnosis
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