Noninvasive Detection of Bladder Cancer via Urinary DNA Mutation and Methylation Profiling Using Modified Real-Time Quantitative PCR

Cystoscopy is the gold standard for bladder cancer (BC) diagnosis and surveillance but experiences invasiveness and patient discomfort. Existing urine-based assays with single-gene mutation or methylation biomarkers exhibit insufficient diagnostic efficacy, necessitating a reliable noninvasive detection approach. This study developed EpiMut-BC, a high-specificity urinary PCR assay integrating DNA mutation and methylation signatures based on pyrophosphorolysis-activated polymerization real-time quantitative PCR with engineered polymerases and blocked primers.