Performance Evaluation of Urine DNA Methylation Testing for the Detection of Urothelial Carcinoma in Patients With Hematuria

Urothelial carcinoma (UC) is the most common malignancy of the urinary system, and early detection and diagnosis are crucial for patient prognosis. Hematuria is a significant clinical manifestation of UC, but its diagnosis often relies on invasive procedures and imaging, posing a substantial burden on patients. With the advancement of molecular biology techniques, urine DNA methylation testing has emerged as a non-invasive and highly sensitive method for early UC detection. This study aims to evaluate the performance of urine DNA methylation testing in detecting UC in patients with hematuria.

All patients with hematuria will receive standard clinical diagnosis, including examinations, tests, surgeries, and pathological assessments to determine the presence of UC. These clinical diagnoses will serve as the gold standard to evaluate the sensitivity and specificity of urine DNA methylation testing in detecting UC in patients with hematuria.

Based on the expected performance indicators of urine DNA methylation testing and previous study data, the sample size calculation is as follows:

Expected sensitivity: 0.86 Hypothesized sensitivity: 0.80 Expected specificity: 0.90 Hypothesized specificity: 0.85 Significance level (Alpha): 0.05 Considering a dropout rate of 5%, the actual number of participants to be recruited is 1109, ensuring an effective sample size of 1053.

Study Procedure Enrollment and Sample Collection: Screen patients meeting the inclusion criteria, obtain informed consent, collect urine samples, and assign screening numbers. Authorized researchers will perform blinding and aliquoting of the enrolled samples to ensure objectivity and impartiality in the testing process. After blinding, perform urine DNA methylation testing according to the test reagent instructions.

To ensure accuracy and reliability of the results, select a subset of samples with known test results for external methylation sequencing. Authorized researchers will unblind the samples, and the statistical team will analyze the results, calculating sensitivity, specificity, positive predictive value, and negative predictive value of the urine DNA methylation test.

Follow-up:

For UC patients: Evaluate the relationship between preoperative urine DNA methylation levels and postoperative recurrence-free survival, progression-free survival, and overall prognosis.

Clinical Trial Report Each center will compile a clinical summary and submit it to the main center. The main center will consolidate the reports from all centers, draft, and publish the final clinical trial report.

It is anticipated that urine DNA methylation testing will demonstrate high sensitivity and specificity as a non-invasive diagnostic method for detecting urothelial carcinoma (UC) in patients with hematuria. This approach can provide clinicians with crucial reference information, reduce the diagnostic burden on patients, and enhance early detection rates and patient prognosis for UC.

Inclusion criteria

• Aged between 18 and 99 years, with gross or microscopic hematuria (RBC ≥ 3 /HPF or RBC > 18 /μL for men, > 33 /μL for women).
• Able to provide 50ml urine for testing before surgery.
• Consent to participate in the study and sign the informed consent form.

Exclusion criteria

• With history of any malignancies (including UC).
• Severe urinary tract infection leading to sepsis.
• Patients with indwelling catheters, nephrostomy, or cystostomy.
• Severe liver or kidney failure or other conditions deemed unsuitable for the study.
• Patients who did not undergo surgical treatment for various reasons.
• Samples with insufficient DNA content or other quality control failures.
• Incomplete clinical or pathological data.
• Patients currently undergoing intravesical or systemic chemotherapy, radiotherapy, immunotherapy, or targeted therapy.