Ignite Creation Date:
2024-06-16 @ 11:48 AM
Last Modification Date:
2024-10-26 @ 3:30 PM
Study NCT ID:
NCT06427993
Status:
NOT_YET_RECRUITING
Last Update Posted:
2024-06-20
First Post:
2024-05-20
Brief Title:
Urine DNA Methylation Detection for Hematuria Evaluation
Sponsor:
Changhai Hospital
Organization:
Changhai Hospital
Study Overview
Official Title:
Urine Exfoliated Cell DNA Methylation Detection for Urothelial Carcinomas Diagnosis in Patients With Hematuria--A Prospective Single-center Cohort Study
Status:
NOT_YET_RECRUITING
Status Verified Date:
2024-05
Last Known Status:
None
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
None
Brief Summary:
Background Hematuria a common symptom of urinary system diseases can result from various causes including infection stones trauma and tumors Urothelial carcinoma UC the most common malignancy of the urinary system often presents with hematuria Current diagnostic methods like urine cytology and cystoscopy have limitations in sensitivity and specificity and cystoscopy is invasive DNA methylation biomarkers offer potential for non-invasive UC detection improving diagnostic accuracy in hematuria patients
Objective This study aims to evaluate the diagnostic performance of DNA methylation biomarkers in detecting UC in patients with hematuria
Methods This prospective pilot study will involve collecting preoperative urine samples from hematuria patients for DNA methylation testing using MSRE-qPCR Sample size calculation was based on an assumed 25 prevalence of UC in hematuria patients resulting in a total of 71 participants after accounting for a 20 dropout rate Sensitivity specificity and diagnostic performance will be assessed using ROC curves
Conclusion This study seeks to validate the effectiveness of urine DNA methylation testing for UC detection in hematuria patients providing a basis for its clinical application and informing the design of larger future studies
Objective This study aims to evaluate the diagnostic performance of DNA methylation biomarkers in detecting UC in patients with hematuria
Methods This prospective pilot study will involve collecting preoperative urine samples from hematuria patients for DNA methylation testing using MSRE-qPCR Sample size calculation was based on an assumed 25 prevalence of UC in hematuria patients resulting in a total of 71 participants after accounting for a 20 dropout rate Sensitivity specificity and diagnostic performance will be assessed using ROC curves
Conclusion This study seeks to validate the effectiveness of urine DNA methylation testing for UC detection in hematuria patients providing a basis for its clinical application and informing the design of larger future studies
Detailed Description:
1 Background Hematuria is a common symptom of urinary system diseases and can be caused by various factors including infection stones trauma and tumors Urothelial carcinoma UC is the most common malignant tumor of the urinary system primarily occurring in the bladder renal pelvis and ureter The early symptoms of UC are often not obvious with hematuria being the most common symptom However current diagnostic methods such as urine cytology and cystoscopy have limitations in sensitivity and specificity for diagnosing UC Additionally cystoscopy is invasive and can cause discomfort for patients DNA methylation biomarkers have shown potential in detecting UC providing a non-invasive method to improve diagnostic sensitivity and specificity especially in patients with hematuria
2 Objective This study aims to evaluate the diagnostic performance of DNA methylation biomarkers in detecting urothelial carcinoma in patients with hematuria By collecting preoperative urine samples from a small cohort of hematuria patients and performing DNA methylation testing we aim to explore the feasibility and advantages of this method in clinical applications
3 Methods
Study Design
This is a prospective pilot study aimed at evaluating the effectiveness of DNA methylation biomarkers in detecting urothelial carcinoma in patients with hematuria
Sample Size Calculation
Based on our previous observations the incidence of urothelial carcinoma in patients with hematuria is 20-30 slightly higher than reported in other literature Therefore we hypothesize that 25 of the patients in the hematuria cohort have UC The group allocation ratio R N-N 7525 3
Assumptions
Area under the curve AUC under H0 05 AUC under H1 08 Power 095 Significance level Alpha 005 Type of data Continuous FPR range 000 to 100
Results Using PASS 150 software for sample size calculation to ensure sufficient statistical power The calculated sample size is
N number of patients with UC 14 N- number of patients without UC 42 Total sample size N 56
Considering a 20 dropout rate the adjusted sample size is
N 18 N- 53 Total sample size N 71
Inclusion and Exclusion Criteria
Inclusion Criteria
Aged between 18 and 99 years with gross or microscopic hematuria 3HP 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 malignancy or concomitant malignancies other than 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
Sample Collection
Clinicians will collect fresh urine samples from enrolled hematuria patients and record their basic information clinical information and medical history
Samples will be randomly numbered and provided to DNA methylation testing personnel to ensure blinding
Testing Method
Urine samples were collected in EP Genomic DNA Kit with an automated nucleic acid extraction instrument Subsequently 100 ng of genomic DNA was used for methylation-sensitive restriction enzyme qPCR MSRE-qPCR detection as described previously Different from bisulfite PCR relying on bisulfite conversion MSRE-qPCR is based on the selective digestion of DNA by methylation-sensitive enzyme followed by qPCR with primers that surround the cutting site
Unblinding and Data Organization
After the last sample is successfully enrolled and tested non-recruiting personnel and testing personnel will unblind the samples and organize clinical and pathological information
Data Analysis
Using pathological results as the gold standard Calculate the sensitivity specificity positive predictive value and negative predictive value of DNA methylation biomarkers
Use statistical methods such as ROC curves to evaluate the diagnostic performance of DNA methylation biomarkers and calculate the AUROC
Ethics and Informed Consent
This study has been approved by the hospitals ethics committee and all participants must sign an informed consent form
4 Conclusion This study aims to validate the effectiveness of urine DNA methylation testing in detecting urothelial carcinoma in patients with hematuria and provide evidence for its clinical application The results of this preliminary study will offer essential data support and design optimization suggestions for future larger-scale studies
2 Objective This study aims to evaluate the diagnostic performance of DNA methylation biomarkers in detecting urothelial carcinoma in patients with hematuria By collecting preoperative urine samples from a small cohort of hematuria patients and performing DNA methylation testing we aim to explore the feasibility and advantages of this method in clinical applications
3 Methods
Study Design
This is a prospective pilot study aimed at evaluating the effectiveness of DNA methylation biomarkers in detecting urothelial carcinoma in patients with hematuria
Sample Size Calculation
Based on our previous observations the incidence of urothelial carcinoma in patients with hematuria is 20-30 slightly higher than reported in other literature Therefore we hypothesize that 25 of the patients in the hematuria cohort have UC The group allocation ratio R N-N 7525 3
Assumptions
Area under the curve AUC under H0 05 AUC under H1 08 Power 095 Significance level Alpha 005 Type of data Continuous FPR range 000 to 100
Results Using PASS 150 software for sample size calculation to ensure sufficient statistical power The calculated sample size is
N number of patients with UC 14 N- number of patients without UC 42 Total sample size N 56
Considering a 20 dropout rate the adjusted sample size is
N 18 N- 53 Total sample size N 71
Inclusion and Exclusion Criteria
Inclusion Criteria
Aged between 18 and 99 years with gross or microscopic hematuria 3HP 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 malignancy or concomitant malignancies other than 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
Sample Collection
Clinicians will collect fresh urine samples from enrolled hematuria patients and record their basic information clinical information and medical history
Samples will be randomly numbered and provided to DNA methylation testing personnel to ensure blinding
Testing Method
Urine samples were collected in EP Genomic DNA Kit with an automated nucleic acid extraction instrument Subsequently 100 ng of genomic DNA was used for methylation-sensitive restriction enzyme qPCR MSRE-qPCR detection as described previously Different from bisulfite PCR relying on bisulfite conversion MSRE-qPCR is based on the selective digestion of DNA by methylation-sensitive enzyme followed by qPCR with primers that surround the cutting site
Unblinding and Data Organization
After the last sample is successfully enrolled and tested non-recruiting personnel and testing personnel will unblind the samples and organize clinical and pathological information
Data Analysis
Using pathological results as the gold standard Calculate the sensitivity specificity positive predictive value and negative predictive value of DNA methylation biomarkers
Use statistical methods such as ROC curves to evaluate the diagnostic performance of DNA methylation biomarkers and calculate the AUROC
Ethics and Informed Consent
This study has been approved by the hospitals ethics committee and all participants must sign an informed consent form
4 Conclusion This study aims to validate the effectiveness of urine DNA methylation testing in detecting urothelial carcinoma in patients with hematuria and provide evidence for its clinical application The results of this preliminary study will offer essential data support and design optimization suggestions for future larger-scale studies
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
False
Is a FDA Regulated Device?:
False
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None