Sampling, Logistics, and Analytics of Urine for RT-qPCR-based Diagnostics

Area of Science:

• Oncology
• Molecular Biology
• Biochemistry

Background:

• Liquid biopsy offers a minimally invasive approach for cancer diagnosis using body fluids like urine.
• RNA biomarkers in urine can reflect gene expression status in tumors, aiding in cancer monitoring.
• Current methods for urine sample analysis require standardized protocols for reliable biomarker detection.

Purpose of the Study:

• To develop and validate a comprehensive protocol for the robust acquisition and analysis of RNA-based tumor markers in urine.
• To ensure the stability and integrity of RNA in urine samples during collection, transport, and storage.
• To establish a reliable method for quantitative detection of RNA biomarkers for bladder cancer diagnosis.

Main Methods:

• Detailed protocol for urine sample collection, stabilization, transportation, and storage.
• Quantitative Polymerase Chain Reaction (qPCR)-based technology for RNA marker analysis.
• Specific primer design and amplicon evaluation for accurate gene assignment.

Main Results:

• The developed protocol ensures RNA stability in urine samples for over 10 days at ambient temperatures.
• The method allows for specific assignment of PCR products to human genomic and transcriptomic data.
• Demonstrated robustness for quantitative detection of RNA-based tumor markers in bladder cancer patients.

Conclusions:

• A validated technical option for robust urine sample acquisition and RNA-based tumor marker detection in bladder cancer is presented.
• The protocol is adaptable for analyzing any RNA-based tumor marker in urine from cancer patients.
• This standardized approach enhances the utility of urine liquid biopsy for cancer diagnostics.