Ultrasensitive circulating tumor DNA analysis enables precision medicine: experimental workflow considerations

Area of Science:

• Oncology
• Molecular Diagnostics
• Biomarker Research

Background:

• Circulating tumor DNA (ctDNA) is a key biomarker for cancer management via liquid biopsies.
• Applications span screening, diagnostics, treatment monitoring, and minimal residual disease detection.
• Significant biological and technical hurdles impede widespread clinical adoption of ctDNA analysis.

Purpose of the Study:

• To review the biological and technical challenges in ctDNA analysis.
• To discuss how preanalytical and analytical methods impact cancer applications.
• To highlight areas for improvement for clinical implementation.

Main Methods:

• Literature review of ctDNA analysis workflows.
• Discussion of preanalytical (sampling) and analytical (data interpretation) factors.
• Examination of challenges affecting various cancer applications.

Main Results:

• ctDNA analysis requires high sensitivity, often needing detection of single DNA molecules.
• The entire workflow, from sampling to data interpretation, needs optimization.
• Current clinical utility reports are limited despite demonstrated potential.

Conclusions:

• Sensitivity remains a key challenge for ctDNA analysis in liquid biopsies.
• Combining mutation analysis with other analytes and improving standardization are potential solutions.
• Enhanced guidelines and standardization are essential for routine clinical use of ctDNA.