Liquid Biopsy for Advanced Cancer: An Amplicon-Based Massively Parallel Sequencing Panel Approach to Precision...

Area of Science:

• Molecular Biology
• Clinical Diagnostics
• Genomics

Background:

• Cell-free DNA (cfDNA) has potential clinical applications, but detecting circulating tumor DNA (ctDNA) faces pre-analytical, analytical, and post-analytical challenges.
• Implementing ctDNA testing in academic labs requires cost-effective and rapid methods for a self-sustaining system.
• Assays must demonstrate clinical utility and adaptability to evolving genomic technologies.

Purpose of the Study:

• To describe a practical and widely applicable massively parallel sequencing (MPS) method for ctDNA mutation testing.
• To address the challenges of implementing ctDNA analysis in a clinical laboratory setting.
• To enhance the sensitivity and specificity of ctDNA detection.

Main Methods:

• Massively parallel sequencing (MPS) approach for ctDNA mutation detection.
• Utilized unique molecular identification (UID) tagging.
• Employed deep sequencing techniques.

Main Results:

• The described MPS method is widely applicable and relatively easy to perform.
• Unique molecular identification tagging and deep sequencing enhance assay sensitivity and specificity.
• The approach facilitates the development of a self-supporting ctDNA testing system.

Conclusions:

• Massively parallel sequencing (MPS) offers a viable and adaptable method for clinical ctDNA mutation testing.
• UID tagging and deep sequencing improve the accuracy of ctDNA detection.
• This approach supports the integration of ctDNA analysis into academic clinical laboratories.