Evaluation of Sensitive Urine DNA-based PENK Methylation Test for Detecting Bladder Cancer in Patients with Hematuria.

Hematuria is a prevalent symptom associated with bladder cancer (BC). However, the invasiveness and cost of cystoscopy, the current gold standard for BC diagnosis in hematuria patients, necessitate the development of a sensitive and accurate non-invasive test. This study introduces and validates a highly sensitive urine-based DNA methylation test. The test improves sensitivity in detecting PENK methylation in urine DNA using Linear Target Enrichment (LTE) followed by quantitative methylation-specific PCR (qMSP) (mePENK-LTE/qMSP). In a case-control study comprising 175 BC patients and 143 non-BC patients with hematuria, the test's optimal cutoff value was determined by distinguishing between two groups, achieved an overall sensitivity of 86.9% and specificity of 91.6%, with an area under the curve (AUC) of 0.892. A prospective validation clinical study involving 366 hematuria patients scheduled for cystoscopy assessed the test's performance. The test demonstrated an overall sensitivity of 84.2% in detecting 38 cases of BC, a specificity of 95.7%, and an AUC of 0.900. Notably, the sensitivity for detecting Ta HG and higher stages of BC reached 92.3%. The test's negative predictive value was 98.2%, and the positive predictive value was 68.7%. These findings highlight the potential of the mePENK-LTE/qMSP test in urine as a promising molecular diagnostic tool for detecting primary BC in hematuria patients, which may reduce the need for cystoscopy.