Diagnostic performance analysis of two-gene methylation in urothelial carcinoma based on small-volume urine sample.

Abstract

Early detection of urothelial carcinoma (UCa) remains challenging because of the limitations of current diagnostic methods. The objective of this study was to evaluate a novel, urine-based H4C6/TWIST1 methylation assay for its diagnostic accuracy in UCa. In a prospective, multicenter study, 743 urine samples (387 patients with UCa and 356 controls) from four hospitals were analyzed. DNA was extracted from 3.5 mL of morning urine, and H4C6/TWIST1 methylation status was assessed by using quantitative polymerase chain reaction analysis. Histopathology served as the reference standard. The assay demonstrated high diagnostic accuracy, with 93.02% sensitivity (95% confidence interval, 90.01%-95.35%) and 92.13% specificity (95% confidence interval, 88.83%-94.71%). The positive and negative predictive values were 92.78% and 92.39%, respectively. Performance was particularly strong for high-grade tumors (97.64%) and muscle-invasive tumors (97.50%). Diagnostic efficacy was consistent across age, sex, and tumor location, covering both lower and upper urinary tract carcinomas. H4C6/TWIST1 methylation detection is a noninvasive, highly accurate, and consistent tool with significant clinical potential for the initial diagnosis and monitoring of UCa across all grades and stages.