Related Experiment Video
Updated: May 6, 2026
06:53
Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids
Published on: June 8, 2019
9.9K
Circulating Tumor DNA Monitoring in Patients with Uveal Melanoma Using Mutation-Agnostic Multiplex Drop-Off ddPCR
Aurore Rampanou1, Manuel Rodrigues2,3, François-Clément Bidard1,3,4
1Circulating Tumor Biomarkers Laboratory, Inserm CIC 2501, Department of Translational Research, Institut Curie, Paris 75005, France.
Analytical Chemistry
|May 4, 2026
Summary
New multiplex assays detect circulating tumor DNA (ctDNA) in uveal melanoma (UM) with high sensitivity. These mutation-agnostic droplet digital PCR (ddPCR) tests offer a cost-effective alternative for monitoring metastatic UM (MUM).
Area of Science:
- Molecular Oncology
- Genetics
- Biotechnology
Background:
- Existing methods for detecting circulating tumor DNA (ctDNA) in uveal melanoma (UM), such as targeted next-generation sequencing (NGS) and simplex droplet digital PCR (ddPCR), have limitations.
- Accurate and sensitive detection of ctDNA is crucial for monitoring metastatic UM (MUM) and guiding treatment decisions.
Purpose of the Study:
- To develop and validate novel mutation-agnostic multiplex drop-off ddPCR assays for enhanced ctDNA detection in UM.
- To evaluate the analytical sensitivity, specificity, and clinical performance of these multiplex assays compared to existing methods.
Main Methods:
- Development of two multiplex drop-off ddPCR assays targeting hotspot mutations in key UM genes (GNAQ, GNA11, SF3B1, PLCB4, CYSLTR2).
- Analytical validation of sensitivity and specificity.
- Clinical validation using tumor and plasma DNA from metastatic UM patients, comparing results with NGS and simplex ddPCR.
Main Results:
- Multiplex assays demonstrated high sensitivity (0.06%-0.13% detection limits) and specificity, comparable to simplex ddPCR.
- Somatic mutations were accurately identified in tumor DNA, and ctDNA was detected in 62.8% of plasma samples from MUM patients.
- Strong concordance was observed between multiplex ddPCR assays and simplex ddPCR, confirming assay accuracy.
Conclusions:
- Mutation-agnostic multiplex drop-off ddPCR assays offer a sensitive, specific, and cost-effective strategy for ctDNA monitoring in UM.
- These assays overcome the genotype-dependency of NGS, enabling broader clinical applicability for real-time treatment monitoring in UM patients.
- The developed assays represent a significant advancement for managing metastatic uveal melanoma.
