Urine Methylation Assay Detects Urothelial Cancer

URINE methylation testing for urothelial carcinoma (UC) achieved 90.8% sensitivity and 93.2% specificity in a multicentre diagnostic study, suggesting potential for non-invasive cancer detection and postoperative surveillance. 

The prospective, blinded study was conducted across three tertiary medical centres in China, with samples collected between December 2021 and June 2025. A total of 2,307 participants were included across a single-centre training cohort, a three-centre validation cohort, and a prospective surveillance cohort. The assay used quantitative methylation-specific polymerase chain reaction to assess urinary DNA methylation targeting AL021918.2 and vimentin (VIM). 

Sensitivity was 91.2% in the training cohort and 90.8% in the validation cohort, while specificity was 90.0% and 93.2%, respectively. Performance remained high for Ta UC and low-grade UC, with validation cohort sensitivities of 84.3% and 82.5%, respectively. 

Methylation Testing Outperformed Cytology for Sensitivity 

The urine methylation assay demonstrated higher sensitivity than urine cytology and fluorescence in situ hybridisation (FISH), while maintaining comparable specificity. This performance suggests that urinary methylation analysis could complement existing approaches to UC detection. 

The study also assessed changes following surgery. Among 32 patients with positive preoperative methylation results, 30 had negative results after surgery, corresponding to 93.8%. 

These findings are relevant to postoperative monitoring because UC is characterised by frequent recurrence and there is an ongoing need for accurate non-invasive surveillance methods. 

Recurrences Detected Before Clinical Confirmation 

During prospective surveillance, the assay detected all nine observed recurrences. The median lead time before clinical confirmation was 4.5 months, with an interquartile range of 1.5-5.5 months. Among 35 patients without recurrence, methylation results remained negative in 29, corresponding to 82.9%. 

The authors concluded that the non-invasive assay showed high diagnostic accuracy and detected all observed recurrences during surveillance. They suggested that urine methylation testing could serve as an adjunct for UC diagnosis and postoperative surveillance, although further evaluation will be needed to establish its role in clinical practice. 

Reference 

Wu Y et al. Urine Methylation test for urothelial cancer diagnosis and surveillance. JAMA Netw Open. 2026;9;(9):e2636786.